Methods for fabricating isolated micro- or nano-structures using soft or imprint lithography

ABSTRACT

The presently disclosed subject matter describes the use of fluorinated elastomer-based materials, in particular perfluoropolyether (PFPE)-based materials, in high-resolution soft or imprint lithographic applications, such as micro- and nanoscale replica molding, and the first nano-contact molding of organic materials to generate high fidelity features using an elastomeric mold. Accordingly, the presently disclosed subject matter describes a method for producing free-standing, isolated nanostructures of any shape using soft or imprint lithography technique.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of pending U.S. patent applicationSer. No. 16/689,733 filed Nov. 20, 2019, which is a continuation of U.S.patent application Ser. No. 15/846,827 filed Dec. 19, 2017, now U.S.Pat. No. 10,517,824 issued on Dec. 31, 2019, which is a continuation ofU.S. patent application Ser. No. 14/658,386 filed Mar. 16, 2015 now U.S.Pat. No. 9,877,920 issued on Jan. 30, 2018, which is a continuation ofU.S. patent application Ser. No. 13/852,683, filed on Mar. 28, 2013, nowU.S. Pat. No. 8,992,992 issued on Mar. 31, 2015, which is a continuationof U.S. patent application Ser. No. 11/825,469, filed on Jul. 6, 2007,now U.S. Pat. No. 8,420,124 issued on Apr. 16, 2013, which is acontinuation of U.S. patent application Ser. No. 10/583,570, filed onMar. 5, 2007, now U.S. Pat. No. 8,263,129 issued on Sep. 11, 2012, whichis a 371 application of PCT International Patent Application Serial No.PCT/US04/042706, filed on Dec. 20, 2004, the disclosure of each of whichis incorporated herein by reference in its entirety, which claimspriority to U.S. Provisional Patent Application Ser. No. 60/531,531,filed Dec. 19, 2003, U.S. Provisional Patent Application Ser. No.60/583,170, filed Jun. 25, 2004, and U.S. Provisional Patent ApplicationSer. No. 60/604,970, filed Aug. 27, 2004, each of which is incorporatedherein by reference in its entirety.

GOVERNMENT INTEREST

This invention was made with U.S. Government support from the Office ofNaval Research Grant No. N00014-02-1-0185 and the Science and TechnologyCenter program of the National Science Foundation under Agreement No.CHE-9876674. The U.S. Government has certain rights in the invention.

TECHNICAL FIELD

Methods for preparing micro- and/or nanoscale particles using soft orimprint lithography. A method for delivering a therapeutic agent to atarget. Methods for forming a micro- or nano-scale pattern on asubstrate using soft or imprint lithography.

Abbreviations

-   -   ° C.=degrees Celsius    -   cm=centimeter    -   DBTDA=dibutyltin diacetate    -   DMA=dimethylacrylate    -   DMPA=2,2-dimethoxy-2-phenylacetophenone    -   EIM=2-isocyanatoethyl methacrylate    -   FEP=fluorinated ethylene propylene    -   Freon 113=1,1,2-trichlorotrifluoroethane    -   g=grams    -   h=hours    -   Hz=hertz    -   IL=imprint lithography    -   kg=kilograms    -   kHz=kilohertz    -   kPa=kilopascal    -   MCP=microcontact printing    -   MEMS=micro-electro-mechanical system    -   MHz=megahertz    -   MIMIC=micro-molding in capillaries    -   mL=milliliters    -   mm=millimeters    -   mmol=millimoles    -   mN=milli-Newton    -   m.p.=melting point    -   mW=milliwatts    -   NCM=nano-contact molding    -   NIL=nanoimprint lithography    -   nm=nanometers    -   PDMS=polydimethylsiloxane    -   PEG poly(ethylene glycol)    -   PFPE=perfluoropolyether    -   PLA poly(lactic acid)    -   PP=polypropylene    -   Ppy=poly(pyrrole)    -   psi=pounds per square inch    -   PVDF=poly(vinylidene fluoride)    -   PTFE=polytetrafluoroethylene    -   SAMIM=solvent-assisted micro-molding    -   SEM=scanning electron microscopy    -   S-FIL=“step and flash” imprint lithography    -   Si=silicon    -   TMPTA=trimethylopropane triacrylate    -   μm=micrometers    -   UV=ultraviolet    -   W=watts    -   ZDOL=poly(tetrafluoroethylene oxide-co-difluoromethylene        oxide)α,ω diol

BACKGROUND

The availability of viable nanofabrication processes is a key factor torealizing the potential of nanotechnologies. In particular, theavailability of viable nanofabrication processes is important to thefields of photonics, electronics, and proteomics. Traditional imprintlithographic (IL) techniques are an alternative to photolithography formanufacturing integrated circuits, micro- and nano-fluidic devices, andother devices with micrometer and/or nanometer sized features. There isa need in the art, however, for new materials to advance IL techniques.See Xia, Y., et al., Angew. Chem. Int. Ed., 1998, 37, 550-575; Xia, Y.,et al., Chem. Rev., 1999, 99, 1823-1848; Resnick, D. J., et al.,Semiconductor International, 2002, June, 71-78; Choi, K. M., et al., J.Am. Chem. Soc., 2003, 125, 4060-4061; McClelland, G. M., et al., Appl.Phys. Lett., 2002, 81, 1483; Chou, S. Y., et al., J. Vac. Sci. Technol.B, 1996, 14, 4129; Otto, M., et al., Microelectron. Eng., 2001, 57, 361;and Bailey, T., et al., J. Vac. Sci. Technol., B, 2000, 18, 3571.

Imprint lithography comprises at least two areas: (1) soft lithographictechniques, see Xia, Y., et al., Angew. Chem. Int. Ed., 1998, 37,550-575, such as solvent-assisted micro-molding (SAMIM); micro-moldingin capillaries (MIMIC); and microcontact printing (MCP); and (2) rigidimprint lithographic techniques, such as nano-contact molding (NCM), seeMcClelland, G. M., et al., Appl. Phys. Lett., 2002, 81, 1483; Otto, M.,et al., Microelectron. Eng., 2001, 57, 361; “step and flash” imprintlithographic (S-FIL), see Bailey, T., et al., J. Vac. Sci. Technol., B,2000, 18, 3571; and nanoimprint lithography (NIL), see Chou, S. Y., etal., J. Vac. Sci. Technol. B, 1996, 14, 4129.

Polydimethylsiloxane (PDMS) based networks have been the material ofchoice for much of the work in soft lithography. See Quake, S. R., etal., Science, 2000, 290, 1536; Y. N. Xia and G. M. Whitesides, Angew.Chem. Int. Ed. Engl. 1998, 37, 551; and Y. N. Xia, et al., Chem. Rev.1999, 99, 1823.

The use of soft, elastomeric materials, such as PDMS, offers severaladvantages for lithographic techniques. For example, PDMS is highlytransparent to ultraviolet (UV) radiation and has a very low Young'smodulus (approximately 750 kPa), which gives it the flexibility requiredfor conformal contact, even over surface irregularities, without thepotential for cracking. In contrast, cracking can occur with molds madefrom brittle, high-modulus materials, such as etched silicon and glass.See Bietsch, A., et al., J. Appl. Phys., 2000, 88, 4310-4318. Further,flexibility in a mold facilitates the easy release of the mold frommasters and replicates without cracking and allows the mold to enduremultiple imprinting steps without damaging fragile features.Additionally, many soft, elastomeric materials are gas permeable, aproperty that can be used to advantage in soft lithography applications.

Although PDMS offers some advantages in soft lithography applications,several properties inherent to PDMS severely limit its capabilities insoft lithography. First, PDMS-based elastomers swell when exposed tomost organic soluble compounds. See Lee, J. N., et al., Anal. Chem.,2003, 75, 6544-6554. Although this property is beneficial inmicrocontact printing (MCP) applications because it allows the mold toadsorb organic inks, see Xia, Y., et al., Angew. Chem. Int. Ed., 1998,37, 550-575, swelling resistance is critically important in the majorityof other soft lithographic techniques, especially for SAMIM and MIMIC,and for IL techniques in which a mold is brought into contact with asmall amount of curable organic monomer or resin. Otherwise, thefidelity of the features on the mold is lost and an unsolvable adhesionproblem ensues due to infiltration of the curable liquid into the mold.Such problems commonly occur with PDMS-based molds because most organicliquids swell PDMS. Organic materials, however, are the materials mostdesirable to mold. Additionally, acidic or basic aqueous solutions reactwith PDMS, causing breakage of the polymer chain.

Secondly, the surface energy of PDMS (approximately 25 mN/m) is not lowenough for soft lithography procedures that require high fidelity. Forthis reason, the patterned surface of PDMS-based molds is oftenfluorinated using a plasma treatment followed by vapor deposition of afluoroalkyl trichlorosilane. See Xia, Y., et al., Angew. Chem. Int. Ed.,1998, 37, 550-575. These fluorine-treated silicones swell, however, whenexposed to organic solvents.

Third, the most commonly-used commercially available form of thematerial used in PDMS molds, e.g., Sylgard 184® (Dow CorningCorporation, Midland, Mich., United States of America) has a modulusthat is too low (approximately 1.5 MPa) for many applications. The lowmodulus of these commonly used PDMS materials results in sagging andbending of features and, as such, is not well suited for processes thatrequire precise pattern placement and alignment. Although researchershave attempted to address this last problem, see Odom, T. W., et al., J.Am. Chem. Soc., 2002, 124, 12112-12113; Odom, T. W. et al., Langmuir,2002, 18, 5314-5320; Schmid, H., et al., Macromolecules, 2000, 33,3042-3049; Csucs, G., et al., Langmuir, 2003, 19, 6104-6109; Trimbach,D., et al., Langmuir, 2003, 19, 10957-10961, the materials chosen stillexhibit poor solvent resistance and require fluorination steps to allowfor the release of the mold.

Rigid materials, such as quartz glass and silicon, also have been usedin imprint lithography. See Xia, Y., et al., Angew. Chem. Int. Ed.,1998, 37, 550-575; Resnick, D. J., et al., Semiconductor International,2002, June, 71-78; McClelland, G. M., et al., Appl. Phys. Lett., 2002,81, 1483; Chou, S. Y., et al., J. Vac. Sci. Technol. B, 1996, 14, 4129;Otto, M., et al., Microelectron. Eng., 2001, 57, 361; and Bailey, T., etal., J. Vac. Sci. Technol., B, 2000, 18, 3571; Chou, S. Y., et al.,Science, 1996, 272, 85-87; Von Werne, T. A., et al., J. Am. Chem. Soc.,2003, 125, 3831-3838; Resnick, D. J., et al., J. Vac. Sci. Technol. B,2003, 21, 2624-2631. These materials are superior to PDMS in modulus andswelling resistance, but lack flexibility. Such lack of flexibilityinhibits conformal contact with the substrate and causes defects in themask and/or replicate during separation.

Another drawback of rigid materials is the necessity to use a costly anddifficult to fabricate hard mold, which is typically made by usingconventional photolithography or electron beam (e-beam) lithography. SeeChou, S. Y., et al., J. Vac. Sci. Technol. B, 1996, 14, 4129. Morerecently, the need to repeatedly use expensive quartz glass or siliconmolds in NCM processes has been eliminated by using an acrylate-basedmold generated from casting a photopolymerizable monomer mixture againsta silicon master. See McClelland, G. M., et al., Appl. Phys. Lett.,2002, 81, 1483, and Jung, G. Y., et al., Nanoletters, 2004, ASAP. Thisapproach also can be limited by swelling of the mold in organicsolvents.

Despite such advances, other disadvantages of fabricating molds fromrigid materials include the necessity to use fluorination steps to lowerthe surface energy of the mold, see Resnick, D. J., et al.,Semiconductor International, 2002, June, 71-78, and the inherent problemof releasing a rigid mold from a rigid substrate without breaking ordamaging the mold or the substrate. See Resnick, D. J., et al.,Semiconductor International, 2002, June, 71-78; Bietsch, A., J. Appl.Phys., 2000, 88, 4310-4318. Khanq, D. Y., et al., Langmuir, 2004, 20,2445-2448, have reported the use of rigid molds composed of thermoformedTeflon AF® (DuPont, Wilmington, Del., United States of America) toaddress the surface energy problem. Fabrication of these molds, however,requires high temperatures and pressures in a melt press, a process thatcould be damaging to the delicate features on a silicon wafer master.Additionally, these molds still exhibit the intrinsic drawbacks of otherrigid materials as outlined hereinabove.

Further, a clear and important limitation of fabricating structures onsemiconductor devices using molds or templates made from hard materialsis the usual formation of a residual or “scum” layer that forms when arigid template is brought into contact with a substrate. Even withelevated applied forces, it is very difficult to completely displaceliquids during this process due to the wetting behavior of the liquidbeing molded, which results in the formation of a scum layer. Thus,there is a need in the art for a method of fabricating a pattern or astructure on a substrate, such as a semiconductor device, which does notresult in the formation of a scum layer.

The fabrication of solvent resistant, microfluidic devices with featureson the order of hundreds of microns from photocurable perfluoropolyether(PFPE) has been reported. See Rolland, J. P., et al., J. Am. Chem. Soc.,2004, 126, 2322-2323. PFPE-based materials are liquids at roomtemperature and can be photochemically cross-linked to yield tough,durable elastomers. Further, PFPE-based materials are highly fluorinatedand resist swelling by organic solvents, such as methylene chloride,tetrahydrofuran, toluene, hexanes, and acetonitrile among others, whichare desirable for use in microchemistry platforms based on elastomericmicrofluidic devices. There is a need in the art, however, to applyPFPE-based materials to the fabrication of nanoscale devices for relatedreasons.

Further, there is a need in the art for improved methods for forming apattern on a substrate, such as method employing a patterned mask. SeeU.S. Pat. No. 4,735,890 to Nakane et al.; U.S. Pat. No. 5,147,763 toKamitakahara et al.; U.S. Pat. No. 5,259,926 to Kuwabara et al.; andInternational PCT Publication No. WO 99/54786 to Jackson et al., each ofwhich is incorporated herein by reference in their entirety.

There also is a need in the art for an improved method for formingisolated structures that can be considered “engineered” structures,including but not limited to particles, shapes, and parts. Usingtraditional IL methods, the scum layer that almost always forms betweenstructures acts to connect or link structures together, thereby makingit difficult, if not impossible to fabricate and/or harvest isolatedstructures.

There also is a need in the art for an improved method for formingmicro- and nanoscale charged particles, in particular polymer electrets.The term “polymer electrets” refers to dielectrics with stored charge,either on the surface or in the bulk, and dielectrics with orienteddipoles, frozen-in, ferrielectric, or ferroelectric. On the macro scale,such materials are used, for example, for electronic packaging andcharge electret devices, such as microphones and the like. See Kressman,R., et al., Space-Charge Electrets, Vol. 2, Laplacian Press, 1999; andHarrison, J. S., et al., Piezoelectic Polymers, NASA/CR-2001-211422,ICASE Report No. 2001-43. Poly(vinylidene fluoride) (PVDF) is oneexample of a polymer electret material. In addition to PVDF, chargeelectret materials, such as polypropylene (PP), Teflon-fluorinatedethylene propylene (FEP), and polytetrafluoroethylene (PTFE), also areconsidered polymer electrets.

Further, there is a need in the art for improved methods for deliveringtherapeutic agents, such as drugs, non-viral gene vectors, DNA, RNA,RNAi, and viral particles, to a target. See Biomedical Polymers,Shalaby, S. W., ed., Harner/Gardner Publications, Inc., Cincinnati,Ohio, 1994; Polymeric Biomaterials, Dumitrin, S., ed., Marcel Dekkar,Inc., New York, N.Y., 1994; Park, K., et al., Biodegradable Hydrogelsfor Drug Delivery, Technomic Publishing Company, Inc., Lancaster, Pa.,1993; Gumarqalieva, et al., Biodegradation and Biodeterioration ofPolymers: Kinetic Aspects, Nova Science Publishers, Inc., Commack, N.Y.,1998; Controlled Drug Delivery, American Chemical Society SymposiumSeries 752, Park, K., and Mrsny, R. J., eds., Washington, D.C., 2000;Cellular Drug Delivery: Principles and Practices, Lu, D. R., and Oie,S., eds., Humana Press, Totowa, N.J., 2004; and Bioreversible Carriersin Drug Design: Theory and Applications, Roche, E. B., ed., PergamonPress, New York, N.Y., 1987. For a description of representativetherapeutic agents for use in such delivery methods, see U.S. Pat. No.6,159,443 to Hallahan, which is incorporated herein by reference in itsentirety.

In sum, there exists a need in the art to identify new materials for usein imprint lithographic techniques. More particularly, there is a needin the art for methods for the fabrication of structures at the tens ofmicron level down to sub-100 nm feature sizes.

SUMMARY

In some embodiments, the presently disclosed subject matter describes amethod for forming one or more particles, the method comprising:

-   -   (a) providing a patterned template and a substrate, wherein the        patterned template comprises a patterned template surface having        a plurality of recessed areas formed therein;    -   (b) disposing a volume of liquid material in or on at least one        of:        -   (i) the patterned template surface; and        -   (ii) the plurality of recessed areas; and    -   (c) forming one or more particles by one of:        -   (i) contacting the patterned template surface with the            substrate and treating the liquid material; and        -   (ii) treating the liquid material.

In some embodiments of the method for forming one or more particles, thepatterned template comprises a solvent resistant, low surface energypolymeric material derived from casting low viscosity liquid materialsonto a master template and then curing the low viscosity liquidmaterials to generate a patterned template. In some embodiments, thepatterned template comprises a solvent resistant elastomeric material.

In some embodiments, at least one of the patterned template andsubstrate comprises a material selected from the group consisting of aperfluoropolyether material, a fluoroolefin material, an acrylatematerial, a silicone material, a styrenic material, a fluorinatedthermoplastic elastomer (TPE), a triazine fluoropolymer, aperfluorocyclobutyl material, a fluorinated epoxy resin, and afluorinated monomer or fluorinated oligomer that can be polymerized orcrosslinked by a metathesis polymerization reaction.

In some embodiments, the presently disclosed subject matter comprises amethod for delivering a therapeutic agent to a target, the methodcomprising:

-   -   (a) providing a particle formed by the method described        hereinabove;    -   (b) admixing the therapeutic agent with the particle; and    -   (c) delivering the particle comprising the therapeutic agent to        the target.        In some embodiments of the method for delivering a therapeutic        agent to a target, the therapeutic agent is selected from one of        a drug and genetic material. In some embodiments, the genetic        material is selected from the group consisting of a non-viral        gene vector, DNA, RNA, RNAi, and a viral particle. In some        embodiments, the particle comprises a biodegradable polymer,        wherein the biodegradable polymer is selected from the group        consisting of a polyester, a polyanhydride, a polyamide, a        phosphorous-based polymer, a poly(cyanoacrylate), a        polyurethane, a polyorthoester, a polydihydropyran, and a        polyacetal.

In some embodiments, the presently disclosed subject matter describes amethod for forming a pattern on a substrate, the method comprising:

-   -   (a) providing a patterned template and a substrate, wherein the        patterned template comprises a patterned template surface having        a plurality of recessed areas formed therein;    -   (b) disposing a volume of liquid material in or on at least one        of:        -   (i) the patterned template surface; and        -   (ii) the plurality of recessed areas;    -   (c) contacting the patterned template surface with the        substrate; and    -   (d) treating the liquid material to form a pattern on the        substrate.

In some embodiments of the method for forming a pattern on a substrate,the patterned template comprises a solvent resistant, low surface energypolymeric material derived from casting low viscosity liquid materialsonto a master template and then curing the low viscosity liquidmaterials to generate a patterned template. In some embodiments, thepatterned template comprises a solvent resistant elastomeric material.

In some embodiments, at least one of the patterned template andsubstrate comprises a material selected from the group consisting of aperfluoropolyether material, a fluoroolefin material, an acrylatematerial, a silicone material, a styrenic material, a fluorinatedthermoplastic elastomer (TPE), a triazine fluoropolymer, aperfluorocyclobutyl material, a fluorinated epoxy resin, and afluorinated monomer or fluorinated oligomer that can be polymerized orcrosslinked by a metathesis polymerization reaction.

Accordingly, it is an object of the present invention to provide a novelmethod of making micro-, nano-, and sub-nanostructures. This and otherobjects are achieved in whole or in part by the presently disclosedsubject matter.

An object of the presently disclosed subject matter having been statedhereinabove, other aspects and objects will become evident as thedescription proceeds when taken in connection with the accompanyingDrawings and Examples as best described herein below.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1D are a schematic representation of an embodiment of thepresently disclosed method for preparing a patterned template.

FIGS. 2A-2F are a schematic representation of the presently disclosedmethod for forming one or more micro- and/or nanoscale particles.

FIGS. 3A-3F are a schematic representation of the presently disclosedmethod for preparing one or more spherical particles.

FIGS. 4A-4D are a schematic representation of the presently disclosedmethod for fabricating charged polymeric particles. FIG. 4A representsthe electrostatic charging of the molded particle during polymerizationor crystallization; FIG. 4B represents a charged nano-disc; FIG. 4Crepresents typical random juxtapositioning of uncharged nano-discs; andFIG. 4D represents the spontaneous aggregation of charged nano-discsinto chain-like structures.

FIGS. 5A-5C are a schematic illustration of multilayer particles thatcan be formed using the presently disclosed soft lithography method.

FIGS. 6A-6C are schematic representation of the presently disclosedmethod for making three-dimensional nanostructures using a softlithography technique.

FIGS. 7A-7F are a schematic representation of an embodiment of thepresently disclosed method for preparing a multi-dimensional complexstructure.

FIGS. 8A-8E are a schematic representation of the presently disclosedimprint lithography process resulting in a “scum layer.”

FIGS. 9A-9E are a schematic representation of the presently disclosedimprint lithography method, which eliminates the “scum layer” by using afunctionalized, non-wetting patterned template and a non-wettingsubstrate.

FIGS. 10A-10E are a schematic representation of the presently disclosedsolvent-assisted micro-molding (SAMIM) method for forming a pattern on asubstrate.

FIG. 11 is a scanning electron micrograph of a silicon master comprising3-μm arrow-shaped patterns.

FIG. 12 is a scanning electron micrograph of a silicon master comprising500 nm conical patterns that are <50 nm at the tip.

FIG. 13 is a scanning electron micrograph of a silicon master comprising200 nm trapezoidal patterns.

FIG. 14 is a scanning electron micrograph of 200-nm isolated trapezoidalparticles of poly(ethylene glycol) (PEG) diacrylate.

FIG. 15 is a scanning electron micrograph of 500-nm isolated conicalparticles of PEG diacrylate.

FIG. 16 is a scanning electron micrograph of 3-μm isolated arrow-shapedparticles of PEG diacrylate.

FIG. 17 is a scanning electron micrograph of 200-nm×750-nm×250-nmrectangular shaped particles of PEG diacrylate.

FIG. 18 is a scanning electron micrograph of 200-nm isolated trapezoidalparticles of trimethylopropane triacrylate (TMPTA).

FIG. 19 is a scanning electron micrograph of 500-nm isolated conicalparticles of TMPTA.

FIG. 20 is a scanning electron micrograph of 500-nm isolated conicalparticles of TMPTA, which have been printed using an embodiment of thepresently described non-wetting imprint lithography method and harvestedmechanically using a doctor blade.

FIG. 21 is a scanning electron micrograph of 200-nm isolated trapezoidalparticles of poly(lactic acid) (PLA).

FIG. 22 is a scanning electron micrograph of 200-nm isolated trapezoidalparticles of poly(lactic acid) (PLA), which have been printed using anembodiment of the presently described non-wetting imprint lithographymethod and harvested mechanically using a doctor blade.

FIG. 23 is a scanning electron micrograph of 3-μm isolated arrow-shapedparticles of PLA.

FIG. 24 is a scanning electron micrograph of 500-nm isolatedconical-shaped particles of PLA.

FIG. 25 is a scanning electron micrograph of 200-nm isolated trapezoidalparticles of poly(pyrrole) (Ppy).

FIG. 26 is a scanning electron micrograph of 3-μm arrow-shaped Ppyparticles.

FIG. 27 is a scanning electron micrograph of 500-nm conical shaped Ppyparticles.

FIGS. 28A-28C are fluorescence confocal micrographs of 200-nm isolatedtrapezoidal particles of PEG diacrylate that contain fluorescentlytagged DNA. FIG. 28A is a fluorescent confocal micrograph of 200 nmtrapezoidal PEG nanoparticles which contain 24-mer DNA strands that aretagged with CY-3. FIG. 28B is optical micrograph of the 200-nm isolatedtrapezoidal particles of PEG diacrylate that contain fluorescentlytagged DNA. FIG. 28C is the overlay of the images provided in FIGS. 28Aand 28B, showing that every particle contains DNA.

FIG. 29 is a scanning electron micrograph of fabrication of 200-nmPEG-diacrylate nanoparticles using “double stamping.”

FIG. 30 is an atomic force micrograph image of 140-nm lines of TMPTAseparated by distance of 70 nm that were fabricated using a PFPE mold.

FIGS. 31A and 31B are a scanning electron micrograph of mold fabricationfrom electron-beam lithographically generated masters. FIG. 31A is ascanning electron micrograph of silicon/silicon oxide masters of 3micron arrows.

FIG. 31B is a scanning electron micrograph of silicon/silicon oxidemasters of 200-nm×800-nm bars.

FIGS. 32A and 32B are an optical micrographic image of mold fabricationfrom photoresist masters. FIG. 32A is a SU-8 master. FIG. 32B is aPFPE-DMA mold templated from a photolithographic master.

FIGS. 33A and 33B are an atomic force micrograph of mold fabricationfrom Tobacco Mosaic Virus templates. FIG. 33A is a master. FIG. 33B is aPFPE-DMA mold templated from a virus master.

FIGS. 34A and 34B are an atomic force micrograph of mold fabricationfrom block copolymer micelle masters. FIG. 34A is apolystyrene-polyisoprene block copolymer micelle. FIG. 34B is a PFPE-DMAmold templated from a micelle master.

FIGS. 35A and 35B are an atomic force micrograph of mold fabricationfrom brush polymer masters. FIG. 35A is a brush polymer master. FIG. 35Bis a PFPE-DMA mold templated from a brush polymer master.

DETAILED DESCRIPTION

The presently disclosed subject matter will now be described more fullyhereinafter with reference to the accompanying Examples, in whichrepresentative embodiments are shown. The presently disclosed subjectmatter can, however, be embodied in different forms and should not beconstrued as limited to the embodiments set forth herein. Rather, theseembodiments are provided so that this disclosure will be thorough andcomplete, and will fully convey the scope of the embodiments to thoseskilled in the art.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this presently described subject matter belongs. Allpublications, patent applications, patents, and other referencesmentioned herein are incorporated by reference in their entirety.

Throughout the specification and claims, a given chemical formula orname shall encompass all optical and stereoisomers, as well as racemicmixtures where such isomers and mixtures exist.

I. Materials

The presently disclosed subject matter broadly describes solventresistant, low surface energy polymeric materials, derived from castinglow viscosity liquid materials onto a master template and then curingthe low viscosity liquid materials to generate a patterned template foruse in high-resolution soft or imprint lithographic applications, suchas micro- and nanoscale replica molding. In some embodiments, thepatterned template comprises a solvent resistant, elastomer-basedmaterial, such as but not limited to a fluorinated elastomer-basedmaterial.

Further, the presently disclosed subject matter describes the firstnano-contact molding of organic materials to generate high fidelityfeatures using an elastomeric mold. Accordingly, the presently disclosedsubject matter describes a method for producing free-standing, isolatedmicro- and nanostructures of any shape using soft or imprint lithographytechniques. Representative micro- and nanostructures include but are notlimited to micro- and nanoparticles, and micro- and nano-patternedsubstrates.

The nanostructures described by the presently disclosed subject mattercan be used in several applications, including, but not limited to,semiconductor manufacturing, such as molding etch barriers without scumlayers for the fabrication of semiconductor devices; crystals; materialsfor displays; photovoltaics; a solar cell device; optoelectronicdevices; routers; gratings; radio frequency identification (RFID)devices; catalysts; fillers and additives; detoxifying agents; etchbarriers; atomic force microscope (AFM) tips; parts for nano-machines;the delivery of a therapeutic agent, such as a drug or genetic material;cosmetics; chemical mechanical planarization (CMP) particles; and porousparticles and shapes of any kind that will enable the nanotechnologyindustry.

Representative solvent resistant elastomer-based materials include butare not limited to fluorinated elastomer-based materials. As usedherein, the term “solvent resistant” refers to a material, such as anelastomeric material that neither swells nor dissolves in commonhydrocarbon-based organic solvents or acidic or basic aqueous solutions.Representative fluorinated elastomer-based materials include but are notlimited to perfluoropolyether (PFPE)-based materials. A photocurableliquid PFPE exhibits desirable properties for soft lithography. Arepresentative scheme for the synthesis and photocuring of functionalPFPEs is provided in Scheme 1.

Additional schemes for the synthesis of functional perfluoropolyethersare provided in Examples 7.1 through 7.6.

This PFPE material has a low surface energy (for example, about 12mN/m); is non-toxic, UV transparent, and highly gas permeable; and curesinto a tough, durable, highly fluorinated elastomer with excellentrelease properties and resistance to swelling. The properties of thesematerials can be tuned over a wide range through the judicious choice ofadditives, fillers, reactive co-monomers, and functionalization agents.Such properties that are desirable to modify, include, but are notlimited to, modulus, tear strength, surface energy, permeability,functionality, mode of cure, solubility and swelling characteristics,and the like. The non-swelling nature and easy release properties of thepresently disclosed PFPE materials allows for nanostructures to befabricated from any material. Further, the presently disclosed subjectmatter can be expanded to large scale rollers or conveyor belttechnology or rapid stamping that allow for the fabrication ofnanostructures on an industrial scale.

In some embodiments, the patterned template comprises a solventresistant, low surface energy polymeric material derived from castinglow viscosity liquid materials onto a master template and then curingthe low viscosity liquid materials to generate a patterned template. Insome embodiments, the patterned template comprises a solvent resistantelastomeric material.

In some embodiments, at least one of the patterned template andsubstrate comprises a material selected from the group consisting of aperfluoropolyether material, a fluoroolefin material, an acrylatematerial, a silicone material, a styrenic material, a fluorinatedthermoplastic elastomer (TPE), a triazine fluoropolymer, aperfluorocyclobutyl material, a fluorinated epoxy resin, and afluorinated monomer or fluorinated oligomer that can be polymerized orcrosslinked by a metathesis polymerization reaction.

In some embodiments, the perfluoropolyether material comprises abackbone structure selected from the group consisting of:

wherein X is present or absent, and when present comprises an endcappinggroup.

In some embodiments, the fluoroolefin material is selected from thegroup consisting of:

wherein CSM comprises a cure site monomer.

In some embodiments, the fluoroolefin material is made from monomerswhich comprise tetrafluoroethylene, vinylidene fluoride,hexafluoropropylene, 2,2-bis(trifluoromethyl)-4,5-difluoro-1,3-dioxole,a functional fluoroolefin, functional acrylic monomer, and a functionalmethacrylic monomer.

In some embodiments, the silicone material comprises a fluoroalkylfunctionalized polydimethylsiloxane (PDMS) having the followingstructure:

wherein:

R is selected from the group consisting of an acrylate, a methacrylate,and a vinyl group; and

Rf comprises a fluoroalkyl chain.

In some embodiments, the styrenic material comprises a fluorinatedstyrene monomer selected from the group consisting of:

wherein Rf comprises a fluoroalkyl chain.

In some embodiments, the acrylate material comprises a fluorinatedacrylate or a fluorinated methacrylate having the following structure:

wherein:

R is selected from the group consisting of H, alkyl, substituted alkyl,aryl, and substituted aryl; and

Rf comprises a fluoroalkyl chain.

In some embodiments, the triazine fluoropolymer comprises a fluorinatedmonomer. In some embodiments, the fluorinated monomer or fluorinatedoligomer that can be polymerized or crosslinked by a metathesispolymerization reaction comprises a functionalized olefin. In someembodiments, the functionalized olefin comprises a functionalized cyclicolefin.

In some embodiments, at least one of the patterned template and thesubstrate has a surface energy lower than 18 mN/m. In some embodiments,at least one of the patterned template and the substrate has a surfaceenergy lower than 15 mN/m.

From a property point of view, the exact properties of these moldingmaterials can be adjusted by adjusting the composition of theingredients used to make the materials. In particular the modulus can beadjusted from low (approximately 1 MPa) to multiple GPa.

II. Formation of Isolated Micro- and/or Nanoparticles

In some embodiments, the presently disclosed subject matter provides amethod for making isolated micro- and/or nanoparticles. In someembodiments, the process comprises initially forming a patternedsubstrate. Turning now to FIG. 1A, a patterned master 100 is provided.Patterned master 100 comprises a plurality of non-recessed surface areas102 and a plurality of recesses 104. In some embodiments, patternedmaster 100 comprises an etched substrate, such as a silicon wafer, whichis etched in the desired pattern to form patterned master 100.

Referring now to FIG. 1B, a liquid material 106, for example, a liquidfluoropolymer composition, such as a PFPE-based precursor, is thenpoured onto patterned master 100. Liquid material 106 is treated bytreating process T_(r), for example exposure to UV light, therebyforming a treated liquid material 108 in the desired pattern.

Referring now to FIGS. 1C and 1D, a force F_(r) is applied to treatedliquid material 108 to remove it from patterned master 100. As shown inFIGS. 1C and 1D, treated liquid material 108 comprises a plurality ofrecesses 110, which are mirror images of the plurality of non-recessedsurface areas 102 of patterned master 100. Continuing with FIGS. 1C and1D, treated liquid material 108 comprises a plurality of first patternedsurface areas 112, which are mirror images of the plurality of recesses104 of patterned master 100. Treated liquid material 108 can now be usedas a patterned template for soft lithography and imprint lithographyapplications. Accordingly, treated liquid material 108 can be used as apatterned template for the formation of isolated micro- andnanoparticles. For the purposes of FIGS. 1A-1D, 2A-2E, and 3A-3F, thenumbering scheme for like structures is retained throughout.

Referring now to FIG. 2A, in some embodiments, a substrate 200, forexample, a silicon wafer, is treated or is coated with a non-wettingmaterial 202. In some embodiments, non-wetting material 202 comprises anelastomer (such a solvent resistant elastomer, including but not limitedto a PFPE elastomer) that can be further exposed to UV light and curedto form a thin, non-wetting layer on the surface of substrate 200.Substrate 200 also can be made non-wetting by treating substrate 200with non-wetting agent 202, for example a small molecule, such as analkyl- or fluoroalkyl-silane, or other surface treatment. Continuingwith FIG. 2A, a droplet 204 of a curable resin, a monomer, or a solutionin which the desired particles will be formed is then placed on thecoated substrate 200.

Referring now to FIG. 2A and FIG. 2B, patterned template 108 (as shownin FIG. 1D) is then contacted with droplet 204 so that droplet 204 fillsthe plurality of recessed areas 110 of patterned template 108.

Referring now to FIGS. 2C and 2D, a force F_(a) is applied to patternedtemplate 108. While not wishing to be bound by any particular theory,once force F_(a) is applied, the affinity of patterned template 108 fornon-wetting coating or surface treatment 202 on substrate 200 incombination with the non-wetting behavior of patterned template 108 andsurface treated or coated substrate 200 causes droplet 204 to beexcluded from all areas except for recessed areas 110. Further, inembodiments essentially free of non-wetting or low wetting material 202with which to sandwich droplet 204, a “scum” layer that interconnectsthe objects being stamped forms.

Continuing with FIGS. 2C and 2D, the material filling recessed areas110, e.g., a resin, monomer, solvent, and combinations thereof, is thentreated by a treating process T_(r), e.g., photocured through patternedtemplate 108 or thermally cured while under pressure, to form aplurality of micro- and/or nanoparticles 206. In some embodiments, amaterial, including but not limited to a polymer, an organic compound,or an inorganic compound, can be dissolved in a solvent, patterned usingpatterned template 108, and the solvent can be released.

Continuing with FIGS. 2C and 2D, once the material filling recessedareas 110 is treated, patterned template 108 is removed from substrate200. Micro- and/or nanoparticles 206 are confined to recessed areas 110of patterned template 108. In some embodiments, micro- and/ornanoparticles 206 can be retained on substrate 200 in defined regionsonce patterned template 108 is removed. This embodiment can be used inthe manufacture of semiconductor devices where essentially scum-layerfree features could be used as etch barriers or as conductive,semiconductive, or dielectric layers directly, mitigating or reducingthe need to use traditional and expensive photolithographic processes.

Referring now to FIGS. 2D and 2E, micro- and/or nanoparticles 206 can beremoved from patterned template 108 to provide freestanding particles bya variety of methods, which include but are not limited to: (1) applyingpatterned template 108 to a surface that has an affinity for theparticles 206; (2) deforming patterned template 108, or using othermechanical methods, including sonication, in such a manner that theparticles 206 are naturally released from patterned template 108; (3)swelling patterned template 108 reversibly with supercritical carbondioxide or another solvent that will extrude the particles 206; and (4)washing patterned template 108 with a solvent that has an affinity forthe particles 206 and will wash them out of patterned template 108.

In some embodiments, the method comprises a batch process. In someembodiments, the batch process is selected from one of a semi-batchprocess and a continuous batch process. Referring now to FIG. 2F, anembodiment of the presently disclosed subject matter wherein particles206 are produced in a continuous process is schematically presented. Anapparatus 199 is provided for carrying out the process. Indeed, whileFIG. 2F schematically presents a continuous process for particles,apparatus 199 can be adapted for batch processes, and for providing apattern on a substrate continuously or in batch, in accordance with thepresently disclosed subject matter and based on a review of thepresently disclosed subject matter by one of ordinary skill in the art.

Continuing, then, with FIG. 2F, droplet 204 of liquid material isapplied to substrate 200′ via reservoir 203. Substrate 200′ can becoated or not coated with a non-wetting agent. Substrate 200′ andpattern template 108′ are placed in a spaced relationship with respectto each other and are also operably disposed with respect to each otherto provide for the conveyance of droplet 204 between patterned template108′ and substrate 200′. Conveyance is facilitated through the provisionof pulleys 208, which are in operative communication with controller201. By way of representative non-limiting examples, controller 201 cancomprise a computing system, appropriate software, a power source, aradiation source, and/or other suitable devices for controlling thefunctions of apparatus 199. Thus, controller 201 provides for power forand other control of the operation of pulleys 208 to provide for theconveyance of droplet 204 between patterned template 108′ and substrate200′. Particles 206 are formed and treated between substrate 200′ andpatterned template 108′ by a treating process T_(R), which is alsocontrolled by controller 201. Particles 206 are collected in aninspecting device 210, which is also controlled by controller 201.Inspecting device 210 provides for one of inspecting, measuring, andboth inspecting and measuring one or more characteristics of particles206. Representative examples of inspecting devices 210 are disclosedelsewhere herein.

Thus, in some embodiments, the method for forming one or more particlescomprises:

-   -   (a) providing a patterned template and a substrate, wherein the        patterned template comprises a first patterned template surface        having a plurality of recessed areas formed therein;    -   (b) disposing a volume of liquid material in or on at least one        of:        -   (i) the first patterned template surface; and        -   (ii) the plurality of recessed areas; and    -   (c) forming one or more particles by one of:        -   (i) contacting the patterned template surface with the            substrate and treating the liquid material; and        -   (ii) treating the liquid material.

In some embodiments of the method for forming one or more particles, thepatterned template comprises a solvent resistant, low surface energypolymeric material derived from casting low viscosity liquid materialsonto a master template and then curing the low viscosity liquidmaterials to generate a patterned template. In some embodiments, thepatterned template comprises a solvent resistant elastomeric material.

In some embodiments, at least one of the patterned template andsubstrate comprises a material selected from the group consisting of aperfluoropolyether material, a fluoroolefin material, an acrylatematerial, a silicone material, a styrenic material, a fluorinatedthermoplastic elastomer (TPE), a triazine fluoropolymer, aperfluorocyclobutyl material, a fluorinated epoxy resin, and afluorinated monomer or fluorinated oligomer that can be polymerized orcrosslinked by a metathesis polymerization reaction.

In some embodiments, the perfluoropolyether material comprises abackbone structure selected from the group consisting of:

wherein X is present or absent, and when present comprises an endcappinggroup.

In some embodiments, the fluoroolefin material is selected from thegroup consisting of:

wherein CSM comprises a cure site monomer.

In some embodiments, the fluoroolefin material is made from monomerswhich comprise tetrafluoroethylene, vinylidene fluoride,hexafluoropropylene, 2,2-bis(trifluoromethyl)-4,5-difluoro-1,3-dioxole,a functional fluoroolefin, functional acrylic monomer, and a functionalmethacrylic monomer.

In some embodiments, the silicone material comprises a fluoroalkylfunctionalized polydimethylsiloxane (PDMS) having the followingstructure:

wherein:

R is selected from the group consisting of an acrylate, a methacrylate,and a vinyl group; and

Rf comprises a fluoroalkyl chain.

In some embodiments, the styrenic material comprises a fluorinatedstyrene monomer selected from the group consisting of:

wherein Rf comprises a fluoroalkyl chain.

In some embodiments, the acrylate material comprises a fluorinatedacrylate or a fluorinated methacrylate having the following structure:

wherein:

R is selected from the group consisting of H, alkyl, substituted alkyl,aryl, and substituted aryl; and

Rf comprises a fluoroalkyl chain.

In some embodiments, the triazine fluoropolymer comprises a fluorinatedmonomer. In some embodiments, the fluorinated monomer or fluorinatedoligomer that can be polymerized or crosslinked by a metathesispolymerization reaction comprises a functionalized olefin. In someembodiments, the functionalized olefin comprises a functionalized cyclicolefin.

In some embodiments, at least one of the patterned template and thesubstrate has a surface energy lower than 18 mN/m. In some embodiments,at least one of the patterned template and the substrate has a surfaceenergy lower than 15 mN/m.

In some embodiments, the substrate is selected from the group consistingof a polymer material, an inorganic material, a silicon material, aquartz material, a glass material, and surface treated variants thereof.In some embodiments, the substrate comprises a patterned area.

In some embodiments, the plurality of recessed areas comprises aplurality of cavities. In some embodiments, the plurality of cavitiescomprises a plurality of structural features. In some embodiments, theplurality of structural features has a dimension ranging from about 10microns to about 1 nanometer in size. In some embodiments, the pluralityof structural features has a dimension ranging from about 10 microns toabout 1 micron in size. In some embodiments, the plurality of structuralfeatures has a dimension ranging from about 1 micron to about 100 nm insize. In some embodiments, the plurality of structural features has adimension ranging from about 100 nm to about 1 nm in size.

In some embodiments, the patterned template comprises a patternedtemplate formed by a replica molding process. In some embodiments, thereplica molding process comprises: providing a master template;contacting a liquid material with the master template; and curing theliquid material to form a patterned template.

In some embodiments, the master template is selected from the groupconsisting of: a template formed from a lithography process; a naturallyoccurring template; and combinations thereof. In some embodiments, thenatural template is selected from one of a biological structure and aself-assembled structure. In some embodiments, the one of a biologicalstructure and a self-assembled structure is selected from the groupconsisting of a naturally occurring crystal, an enzyme, a virus, aprotein, a micelle, and a tissue surface.

In some embodiments, the method comprises modifying the patternedtemplate surface by a surface modification step. In some embodiments,the surface modification step is selected from the group consisting of aplasma treatment, a chemical treatment, and an adsorption process. Insome embodiments, the adsorption process comprises adsorbing moleculesselected from the group consisting of a polyelectrolyte, apoly(vinylalcohol), an alkylhalosilane, and a ligand.

In some embodiments, the method comprises positioning the patternedtemplate and the substrate in a spaced relationship to each other suchthat the patterned template surface and the substrate face each other ina predetermined alignment.

In some embodiments, the liquid material is selected from the groupconsisting of a polymer, a solution, a monomer, a plurality of monomers,a polymerization initiator, a polymerization catalyst, an inorganicprecursor, a metal precursor, a pharmaceutical agent, a tag, a magneticmaterial, a paramagnetic material, a ligand, a cell penetrating peptide,a porogen, a surfactant, a plurality of immiscible liquids, a solvent, acharged species, and combinations thereof.

In some embodiments, the pharmaceutical agent is selected from the groupconsisting of a drug, a peptide, RNAi, and DNA. In some embodiments, thetag is selected from the group consisting of a fluorescence tag, aradiolabeled tag, and a contrast agent. In some embodiments, the ligandcomprises a cell targeting peptide.

In some embodiments, the liquid material comprises a non-wetting agent.In some embodiments, the liquid material comprises one phase. In someembodiments, the liquid material comprises a plurality of phases. Insome embodiments, the liquid material is selected from the groupconsisting of multiple liquids, multiple immiscible liquids,surfactants, dispersions, emulsions, micro-emulsions, micelles,particulates, colloids, porogens, active ingredients, and combinationsthereof.

In some embodiments, the disposing of the volume of liquid material onone of the patterned template and the substrate is regulated by aspreading process. In some embodiments, the spreading process comprises:

-   -   (a) disposing a first volume of liquid material on one of the        patterned template and the substrate to form a layer of liquid        material thereon; and    -   (b) drawing an implement across the layer of liquid material to:        -   (i) remove a second volume of liquid material from the layer            of liquid material on the one of the patterned template and            the substrate; and        -   (ii) leave a third volume of liquid material on the one of            the patterned template and the substrate.

In some embodiments, an article is contacted with the layer of liquidmaterial and a force is applied to the article to thereby remove theliquid material from the one of the patterned material and thesubstrate. In some embodiments, the article is selected from the groupconsisting of a roller and a “squeegee” blade. In some embodiments, theliquid material is removed by some other mechanical means.

In some embodiments, the contacting of the patterned template surfacewith the substrate forces essentially all of the disposed liquidmaterial from between the patterned template surface and the substrate.

In some embodiments, the treating of the liquid material comprises aprocess selected from the group consisting of a thermal process, aphotochemical process, and a chemical process.

In some embodiments as described in detail herein below, the methodfurther comprises:

-   -   (a) reducing the volume of the liquid material disposed in the        plurality of recessed areas by one of:        -   (i) applying a contact pressure to the patterned template            surface; and        -   (ii) allowing a second volume of the liquid to evaporate or            permeate through the template;    -   (b) removing the contact pressure applied to the patterned        template surface;    -   (c) introducing gas within the recessed areas of the patterned        template surface;    -   (d) treating the liquid material to form one or more particles        within the recessed areas of the patterned template surface; and    -   (e) releasing the one or more particles.

In some embodiments, the releasing of the one or more particles isperformed by one of:

-   -   (a) applying the patterned template to a substrate, wherein the        substrate has an affinity for the one or more particles;    -   (b) deforming the patterned template such that the one or more        particles is released from the patterned template;    -   (c) swelling the patterned template with a first solvent to        extrude the one or more particles;    -   (d) washing the patterned template with a second solvent,        wherein the second solvent has an affinity for the one or more        particles; and    -   (e) applying a mechanical force to the one or more particles.        In some embodiments, the mechanical force is applied by        contacting one of a Doctor blade and a brush with the one or        more particles. In some embodiments, the mechanical force is        applied by ultrasonics, megasonics, electrostatics, or magnetics        means.

In some embodiments, the method comprises harvesting or collecting theparticles. In some embodiments, the harvesting or collecting of theparticles comprises a process selected from the group consisting ofscraping with a doctor blade, a brushing process, a dissolution process,an ultrasound process, a megasonics process, an electrostatic process,and a magnetic process.

In some embodiments, the presently disclosed subject matter describes aparticle or plurality of particles formed by the methods describedherein. In some embodiments, the plurality of particles comprises aplurality of monodisperse particles. In some embodiments, the particleor plurality of particles is selected from the group consisting of asemiconductor device, a crystal, a drug delivery vector, a gene deliveryvector, a disease detecting device, a disease locating device, aphotovoltaic device, a porogen, a cosmetic, an electret, an additive, acatalyst, a sensor, a detoxifying agent, an abrasive, such as a CMP, amicro-electro-mechanical system (MEMS), a cellular scaffold, a taggant,a pharmaceutical agent, and a biomarker. In some embodiments, theparticle or plurality of particles comprise a freestanding structure.

Further, in some embodiments, the presently disclosed subject matterdescribes a method of fabricating isolated liquid objects, the methodcomprising (a) contacting a liquid material with the surface of a firstlow surface energy material; (b) contacting the surface of a second lowsurface energy material with the liquid, wherein at least one of thesurfaces of either the first or second low surface energy material ispatterned; (c) sealing the surfaces of the first and the second lowsurface energy materials together; and (d) separating the two lowsurface energy materials to produce a replica pattern comprising liquiddroplets.

In some embodiments, the liquid material comprises poly(ethyleneglycol)-diacrylate. In some embodiments, the low surface energy materialcomprises perfluoropolyether-diacrylate. In some embodiments, a chemicalprocess is used to seal the surfaces of the first and the second lowsurface energy materials. In some embodiments, a physical process isused to seal the surfaces of the first and the second low surface energymaterials. In some embodiments, one of the surfaces of the low surfaceenergy material is patterned. In some embodiments, one of the surfacesof the low surface energy material is not patterned.

In some embodiments, the method further comprises using the replicapattern composed of liquid droplets to fabricate other objects. In someembodiments, the replica pattern of liquid droplets is formed on thesurface of the low surface energy material that is not patterned. Insome embodiments, the liquid droplets undergo direct or partialsolidification. In some embodiments, the liquid droplets undergo achemical transformation. In some embodiments, the solidification of theliquid droplets or the chemical transformation of the liquid dropletsproduce freestanding objects. In some embodiments, the freestandingobjects are harvested. In some embodiments, the freestanding objects arebonded in place. In some embodiments, the freestanding objects aredirectly solidified, partially solidified, or chemically transformed.

In some embodiments, the liquid droplets are directly solidified,partially solidified, or chemically transformed on or in the patternedtemplate to produce objects embedded in the recesses of the patternedtemplate. In some embodiments, the embedded objects are harvested. Insome embodiments, the embedded objects are bonded in place. In someembodiments, the embedded objects are used in other fabricationprocesses.

In some embodiments, the replica pattern of liquid droplets istransferred to other surfaces. In some embodiments, the transfer takesplace before the solidification or chemical transformation process. Insome embodiments, the transfer takes place after the solidification orchemical transformation process. In some embodiments, the surface towhich the replica pattern of liquid droplets is transferred is selectedfrom the group consisting of a non-low surface energy surface, a lowsurface energy surface, a functionalized surface, and a sacrificialsurface. In some embodiments, the method produces a pattern on a surfacethat is essentially free of one or more scum layers. In someembodiments, the method is used to fabricate semiconductors and otherelectronic and photonic devices or arrays. In some embodiments, themethod is used to create freestanding objects. In some embodiments, themethod is used to create three-dimensional objects using multiplepatterning steps. In some embodiments, the isolated or patterned objectcomprises materials selected from the group consisting of organic,inorganic, polymeric, and biological materials. In some embodiments, asurface adhesive agent is used to anchor the isolated structures on asurface.

In some embodiments, the liquid droplet arrays or solid arrays onpatterned or non-patterned surfaces are used as regiospecific deliverydevices or reaction vessels for additional chemical processing steps. Insome embodiments, the additional chemical processing steps are selectedfrom the group consisting of printing of organic, inorganic, polymeric,biological, and catalytic systems onto surfaces; synthesis of organic,inorganic, polymeric, biological materials; and other applications inwhich localized delivery of materials to surfaces is desired.Applications of the presently disclosed subject matter include, but arenot limited to, micro and nanoscale patterning or printing of materials.In some embodiments, the materials to be patterned or printed areselected from the group consisting of surface-binding molecules,inorganic compounds, organic compounds, polymers, biological molecules,nanoparticles, viruses, biological arrays, and the like.

In some embodiments, the applications of the presently disclosed subjectmatter include, but are not limited to, the synthesis of polymerbrushes, catalyst patterning for CVD carbon nanotube growth, cellscaffold fabrication, the application of patterned sacrificial layers,such as etch resists, and the combinatorial fabrication of organic,inorganic, polymeric, and biological arrays.

In some embodiments, non-wetting imprint lithography, and relatedtechniques, are combined with methods to control the location andorientation of chemical components within an individual object. In someembodiments, such methods improve the performance of an object byrationally structuring the object so that it is optimized for aparticular application. In some embodiments, the method comprisesincorporating biological targeting agents into particles for drugdelivery, vaccination, and other applications. In some embodiments, themethod comprises designing the particles to include a specificbiological recognition motif. In some embodiments, the biologicalrecognition motif comprises biotin/avidin and/or other proteins.

In some embodiments, the method comprises tailoring the chemicalcomposition of these materials and controlling the reaction conditions,whereby it is then possible to organize the biorecognition motifs sothat the efficacy of the particle is optimized. In some embodiments, theparticles are designed and synthesized so that recognition elements arelocated on the surface of the particle in such a way to be accessible tocellular binding sites, wherein the core of the particle is preserved tocontain bioactive agents, such as therapeutic molecules. In someembodiments, a non-wetting imprint lithography method is used tofabricate the objects, wherein the objects are optimized for aparticular application by incorporating functional motifs, such asbiorecognition agents, into the object composition. In some embodiments,the method further comprises controlling the microscale and nanoscalestructure of the object by using methods selected from the groupconsisting of self-assembly, stepwise fabrication procedures, reactionconditions, chemical composition, crosslinking, branching, hydrogenbonding, ionic interactions, covalent interactions, and the like. Insome embodiments, the method further comprises controlling themicroscale and nanoscale structure of the object by incorporatingchemically organized precursors into the object. In some embodiments,the chemically organized precursors are selected from the groupconsisting of block copolymers and core-shell structures.

In sum, the presently disclosed subject matter describes a non-wettingimprint lithography technique that is scalable and offers a simple,direct route to such particles without the use of self-assembled,difficult to fabricate block copolymers and other systems.

III. Formation of Rounded Particles Through “Liquid Reduction”

Referring now to FIGS. 3A through 3F, the presently disclosed subjectmatter provides a “liquid reduction” process for forming particles thathave shapes that are not conformal to the shape of the template,including but not limited to spherical micro- and nanoparticles. Forexample, a “cube-shaped” template can allow for sphereical particles tobe made, whereas a “Block arrow-shaped” template can allow for“lolli-pop” shaped particles or objects to be made wherein theintroduction of a gas allows surface tension forces to reshape theresident liquid prior to treating it. While not wishing to be bound byany particular theory, the non-wetting characteristics that can beprovided in some embodiments of the presently disclosed patternedtemplate and/or treated or coated substrate allows for the generation ofrounded, e.g., spherical, particles.

Referring now to FIG. 3A, droplet 302 of a liquid material is disposedon substrate 300, which in some embodiments is coated or treated with anon-wetting material 304. A patterned template 108, which comprises aplurality of recessed areas 110 and patterned surface areas 112, also isprovided.

Referring now to FIG. 3B, patterned template 108 is contacted withdroplet 302. The liquid material comprising droplet 302 then entersrecessed areas 110 of patterned template 108. In some embodiments, aresidual, or “scum,” layer RL of the liquid material comprising droplet302 remains between the patterned template 108 and substrate 300.

Referring now to FIG. 3C, a first force F_(a1) is applied to patternedtemplate 108. A contact point CP is formed between the patternedtemplate 108 and the substrate and displacing residual layer RL.Particles 306 are formed in the recessed areas 110 of patterned template108.

Referring now to FIG. 3D, a second force F_(a2), wherein the forceapplied by F_(a2) is greater than the force applied by F_(a1), is thenapplied to patterned template 108, thereby forming smaller liquidparticles 308 inside recessed areas 112 and forcing a portion of theliquid material comprising droplet 302 out of recessed areas 112.

Referring now to FIG. 3E, the second force F_(a2) is released, therebyreturning the contact pressure to the original contact pressure appliedby first force F_(a1). In some embodiments, patterned template 108comprises a gas permeable material, which allows a portion of space withrecessed areas 112 to be filled with a gas, such as nitrogen, therebyforming a plurality of liquid spherical droplets 310. Once this liquidreduction is achieved, the plurality of liquid spherical droplets 310are treated by a treating process T_(r).

Referring now to FIG. 3F, treated liquid spherical droplets 310 arereleased from patterned template 108 to provide a plurality offreestanding spherical particles 312.

IV. Formation of Polymeric Nano- to Micro-Electrets

Referring now to FIGS. 4A and 4B, in some embodiments, the presentlydisclosed subject matter describes a method for preparing polymericnano- to micro-electrets by applying an electric field during thepolymerization and/or crystallization step during molding (FIG. 4A) toyield a charged polymeric particle (FIG. 4B). In some embodiments, thecharged polymeric particles spontaneously aggregate into chain-likestructures (FIG. 4D) instead of the random configurations shown in FIG.4C.

In some embodiments, the charged polymeric particle comprises apolymeric electret. In some embodiments, the polymeric electretcomprises a polymeric nano-electret. In some embodiments, the chargedpolymeric particles aggregate into chain-like structures. In someembodiments, the charged polymeric particles comprise an additive for anelectro-rheological device. In some embodiments, the electro-rheologicaldevice is selected from the group consisting of clutches and activedampening devices. In some embodiments, the charged polymeric particlescomprise nano-piezoelectric devices. In some embodiments, thenano-piezoelectric devices are selected from the group consisting ofactuators, switches, and mechanical sensors.

V. Formation of Multilayer Structures

In some embodiments, the presently disclosed subject matter provides amethod for forming multilayer structures, including multilayerparticles. In some embodiments, the multilayer structures, includingmultilayer particles, comprise nanoscale multilayer structures. In someembodiments, multilayer structures are formed by depositing multiplethin layers of immisible liquids and/or solutions onto a substrate andforming particles as described by any of the methods hereinabove. Theimmiscibility of the liquid can be based on any physical characteristic,including but not limited to density, polarity, and volatility. Examplesof possible morphologies of the presently disclosed subject matter areillustrated in FIGS. 5A-5C and include, but are not limited to,multi-phase sandwich structures, core-shell particles, and internalemulsions, microemulsions and/or nano-sized emulsions.

Referring now to FIG. 5A, a multi-phase sandwich structure 500 of thepresently disclosed subject matter is shown, which by way of example,comprises a first liquid material 502 and a second liquid material 504.

Referring now to FIG. 5B, a core-shell particle 506 of the presentlydisclosed subject matter is shown, which by way of example, comprises afirst liquid material 502 and a second liquid material 504.

Referring now to FIG. 5C, an internal emulsion particle 508 of thepresently disclosed subject matter is shown, which by way of example,comprises a first liquid material 502 and a second liquid material 504.

More particularly, in some embodiments, the method comprises disposing aplurality of immiscible liquids between the patterned template andsubstrate to form a multilayer structure, e.g., a multilayernanostructure. In some embodiments, the multilayer structure comprises amultilayer particle. In some embodiments, the multilayer structurecomprises a structure selected from the group consisting of multi-phasesandwich structures, core-shell particles, internal emulsions,microemulsions, and nanosized emulsions.

VI. Fabrication of Complex Multi-Dimensional Structures

In some embodiments, the currently disclosed subject matter provides aprocess for fabricating complex, multi-dimensional structures. In someembodiments, complex multi-dimensional structures can be formed byperforming the steps illustrated in FIGS. 2A-2E. In some embodiments,the method comprises imprinting onto a patterned template that isaligned with a second patterned template (instead of imprinting onto asmooth substrate) to generate isolated multi-dimensional structures thatare cured and released as described herein. A schematic illustration ofan embodiment of a process for forming complex multi-dimensionalstructures and examples of such structures are provided in FIGS. 6A-6C.

Referring now to FIG. 6A, a first patterned template 600 is provided.First patterned template 600 comprises a plurality of recessed areas 602and a plurality of non-recessed surfaces 604. Also provided is a secondpatterned template 606. Second patterned template 606 comprises aplurality of recessed areas 608 and a plurality of non-recessed surfaces610. As shown in FIG. 6A, first patterned template 600 and secondpatterned template 606 are aligned in a predetermined spacedrelationship. A droplet of liquid material 612 is disposed between firstpatterned template 600 and second patterned template 606.

Referring now to FIG. 6B, patterned template 600 is contacted withpatterned template 606. A force F_(a) is applied to patterned template600 causing the liquid material comprising droplet 612 to migrate to theplurality of recessed areas 602 and 608. The liquid material comprisingdroplet 612 is then treated by treating process T_(r) to form apatterned, treated liquid material 614.

Referring now to FIG. 6C, the patterned, treated liquid material 614 ofFIG. 6B is released by any of the releasing methods described herein toprovide a plurality of multi-dimensional patterned structures 616.

In some embodiments, patterned structure 616 comprises ananoscale-patterned structure. In some embodiments, patterned structure616 comprises a multi-dimensional structure. In some embodiments, themulti-dimensional structure comprises a nanoscale multi-dimensionalstructure. In some embodiments, the multi-dimensional structurecomprises a plurality of structural features. In some embodiments, thestructural features comprise a plurality of heights.

In some embodiments, a microelectronic device comprising patternedstructure 616 is provided. Indeed, patterned structure 616 can be anystructure imaginable, including “dual damscene” structures formicroelectronics. In some embodiments, the microelectronic device isselected from the group consisting of integrated circuits, semiconductorparticles, quantum dots, and dual damascene structures. In someembodiments, the microelectronic device exhibits certain physicalproperties selected from the group consisting of etch resistance, lowdielectric constant, high dielectric constant, conducting,semiconducting, insulating, porosity, and non-porosity.

In some embodiments, the presently disclosed subject matter discloses amethod of preparing a multidimensional, complex structure. Referring nowto FIGS. 7A-7F, in some embodiments, a first patterned template 700 isprovided. First patterned template 700 comprises a plurality ofnon-recessed surface areas 702 and a plurality of recessed surface areas704. Continuing particularly with FIG. 7A, also provided is a substrate706. In some embodiments, substrate 706 is coated with a non-wettingagent 708. A droplet of a first liquid material 710 is disposed onsubstrate 706.

Referring now to FIGS. 7B and 7C, first patterned template 700 iscontacted with substrate 706. A force F_(a) is applied to firstpatterned template 700 such that the droplet of the first liquidmaterial 710 is forced into recesses 704. The liquid material comprisingthe droplet of first liquid material 710 is treated by a first treatingprocess T_(r1) to form a treated first liquid material within theplurality of recesses 704. In some embodiments, first treating processT_(r1) comprises a partial curing process causing the treated firstliquid material to adhere to substrate 706. Referring particularly toFIG. 7C, first patterned template 700 is removed to provide a pluralityof structural features 712 on substrate 706.

Referring now to FIGS. 7D-7F, a second patterned template 714 isprovided. Second patterned substrate 714 comprises a plurality ofrecesses 716, which are filled with a second liquid material 718. Thefilling of recesses 716 can be accomplished in a manner similar to thatdescribed in FIGS. 7A and 7B with respect to recesses 704. Referringparticularly to FIG. 7E, second patterned template 714 is contacted withstructural features 712. Second liquid material 718 is treated with asecond treating process T_(r2) such that the second liquid material 718adheres to the plurality of structural feature 712, thereby forming amultidimensional structure 720. Referring particularly to FIG. 7F,second patterned template 714 and substrate 706 are removed, providing aplurality of free standing multidimensional structures 722. In someembodiments, the process schematically presented in FIGS. 7A-7F can becarried out multiple times as desired to form intricate nanostructures.

Accordingly, in some embodiments, a method for forming multidimensionalstructures is provided, the method comprising:

(a) providing a particle prepared by the process described in thefigures;

(b) providing a second patterned template;

(c) disposing a second liquid material in the second patterned template;

(d) contacting the second patterned template with the particle of step(a); and

(e) treating the second liquid material to form a multidimensionalstructure.

VII. Imprint Lithography

Referring now to FIGS. 8A-8D, a method for forming a pattern on asubstrate is illustrated. In the embodiment illustrated in FIG. 8 , animprint lithography technique is used to form a pattern on a substrate.

Referring now to FIG. 8A, a patterned template 810 is provided. In someembodiments, patterned template 810 comprises a solvent resistant, lowsurface energy polymeric material, derived from casting low viscosityliquid materials onto a master template and then curing the lowviscosity liquid materials to generate a patterned template as definedhereinabove. Patterned template 810 further comprises a first patternedtemplate surface 812 and a second template surface 814. The firstpatterned template surface 812 further comprises a plurality of recesses816. The patterned template derived from a solvent resistant, lowsurface energy polymeric material could be mounted on another materialto facilitate alignment of the patterned template or to facilitatecontinuous processing such as a conveyor belt. This might beparticularly useful in the fabrication of precisely placed structures ona surface, such as in the fabrication of a complex devices or asemiconductor, electronic or photonic devices.

Referring again to FIG. 8A, a substrate 820 is provided. Substrate 820comprises a substrate surface 822. In some embodiments, substrate 820 isselected from the group consisting of a polymer material, an inorganicmaterial, a silicon material, a quartz material, a glass material, andsurface treated variants thereof. In some embodiments, at least one ofpatterned template 810 and substrate 820 has a surface energy lower than18 mN/m. In some embodiments, at least one of patterned template 810 andsubstrate 820 has a surface energy lower than 15 mN/m.

In some embodiments, as illustrated in FIG. 8A, patterned template 810and substrate 820 are positioned in a spaced relationship to each othersuch that first patterned template surface 812 faces substrate surface822 and a gap 830 is created between first patterned template surface812 and substrate surface 822. This is an example of a predeterminedrelationship.

Referring now to FIG. 8B, a volume of liquid material 840 is disposed inthe gap 830 between first patterned template surface 812 and substratesurface 822. In some embodiments, the volume of liquid material 840 isdisposed directed on a non-wetting agent (not shown), which is disposedon first patterned template surface 812.

Referring now to FIG. 8C, in some embodiments, first patterned template812 is contacted with the volume of liquid material 840. A force F_(a)is applied to second template surface 814 thereby forcing the volume ofliquid material 840 into the plurality of recesses 816. In someembodiments, as illustrated in FIG. 8C, a portion of the volume ofliquid material 840 remains between first patterned template surface 812and substrate surface 820 after force F_(a) is applied.

Referring again to FIG. 8C, in some embodiments, the volume of liquidmaterial 840 is treated by a treating process T_(r) while force F_(a) isbeing applied to form a treated liquid material 842. In someembodiments, treating process T_(r) comprises a process selected fromthe group consisting of a thermal process, a photochemical process, anda chemical process.

Referring now to FIG. 8D, a force F_(r) is applied to patterned template810 to remove patterned template 810 from treated liquid material 842 toreveal a pattern 850 on substrate 820 as shown in FIG. 8E. In someembodiments, a residual, or “scum,” layer 852 of treated liquid material842 remains on substrate 820.

More particularly, the method for forming a pattern on a substratecomprises:

-   -   (a) providing patterned template and a substrate, wherein the        patterned template comprises a patterned template surface having        a plurality of recessed areas formed therein,    -   (b) disposing a volume of liquid material in or on at least one        of:        -   (i) the patterned template surface; and        -   (ii) the plurality of recessed areas;    -   (c) contacting the patterned template surface with the        substrate; and    -   (d) treating the liquid material to form a pattern on the        substrate.

In some embodiments, the patterned template comprises a solventresistant, low surface energy polymeric material derived from castinglow viscosity liquid materials onto a master template and then curingthe low viscosity liquid materials to generate a patterned template. Insome embodiments, the patterned template comprises a solvent resistantelastomeric material.

In some embodiments, at least one of the patterned template andsubstrate comprises a material selected from the group consisting of aperfluoropolyether material, a fluoroolefin material, an acrylatematerial, a silicone material, a styrenic material, a fluorinatedthermoplastic elastomer (TPE), a triazine fluoropolymer, aperfluorocyclobutyl material, a fluorinated epoxy resin, and afluorinated monomer or fluorinated oligomer that can be polymerized orcrosslinked by a metathesis polymerization reaction.

In some embodiments, the perfluoropolyether material comprises abackbone structure selected from the group consisting of:

wherein X is present or absent, and when present comprises an endcappinggroup.

In some embodiments, the fluoroolefin material is selected from thegroup consisting of:

wherein CSM comprises a cure site monomer.

In some embodiments, the fluoroolefin material is made from monomers,which comprise tetrafluoroethylene, vinylidene fluoride,hexafluoropropylene, 2,2-bis(trifluoromethyl)-4,5-difluoro-1,3-dioxole,a functional fluoroolefin, functional acrylic monomer, and a functionalmethacrylic monomer.

In some embodiments, the silicone material comprises a fluoroalkylfunctionalized polydimethylsiloxane (PDMS) having the followingstructure:

wherein:

R is selected from the group consisting of an acrylate, a methacrylate,and a vinyl group; and

Rf comprises a fluoroalkyl chain.

In some embodiments, the styrenic material comprises a fluorinatedstyrene monomer selected from the group consisting of:

wherein Rf comprises a fluoroalkyl chain.

In some embodiments, the acrylate material comprises a fluorinatedacrylate or a fluorinated methacrylate having the following structure:

wherein:

R is selected from the group consisting of H, alkyl, substituted alkyl,aryl, and substituted aryl; and

Rf comprises a fluoroalkyl chain.

In some embodiments, the triazine fluoropolymer comprises a fluorinatedmonomer.

In some embodiments, the fluorinated monomer or fluorinated oligomerthat can be polymerized or crosslinked by a metathesis polymerizationreaction comprises a functionalized olefin. In some embodiments, thefunctionalized olefin comprises a functionalized cyclic olefin.

In some embodiments, at least one of the patterned template and thesubstrate has a surface energy lower than 18 mN/m. In some embodiments,at least one of the patterned template and the substrate has a surfaceenergy lower than 15 mN/m.

In some embodiments, the substrate is selected from the group consistingof a polymer material, an inorganic material, a silicon material, aquartz material, a glass material, and surface treated variants thereof.In some embodiments, the substrate is selected from one of an electronicdevice in the process of being manufactured and a photonic device in theprocess of being manufactured. In some embodiments, the substratecomprises a patterned area.

In some embodiments, the plurality of recessed areas comprises aplurality of cavities. In some embodiments, the plurality of cavitiescomprise a plurality of structural features. In some embodiments, theplurality of structural features has a dimension ranging from about 10microns to about 1 nanometer in size. In some embodiments, the pluralityof structural features has a dimension ranging from about 10 microns toabout 1 micron in size. In some embodiments, the plurality of structuralfeatures has a dimension ranging from about 1 micron to about 100 nm insize. In some embodiments, the plurality of structural features has adimension ranging from about 100 nm to about 1 nm in size.

In some embodiments, the liquid material is selected from the groupconsisting of a polymer, a solution, a monomer, a plurality of monomers,a polymerization initiator, a polymerization catalyst, an inorganicprecursor, a metal precursor, a pharmaceutical agent, a tag, a magneticmaterial, a paramagnetic material, a superparamagnetic material, aligand, a cell penetrating peptide, a porogen, a surfactant, a pluralityof immiscible liquids, a solvent, and a charged species. In someembodiments, the pharmaceutical agent is selected from the groupconsisting of a drug, a peptide, RNAi, and DNA. In some embodiments, thetag is selected from the group consisting of a fluorescence tag, aradiolabeled tag, and a contrast agent. In some embodiments, the ligandcomprises a cell targeting peptide.

Representative superparamagnetic or paramagnetic materials include butare not limited to Fe₂O₃, Fe₃O₄, FePt, Co, MnFe₂O₄, CoFe₂O₄, CuFe₂O₄,NiFe₂O₄ and ZnS doped with Mn for magneto-optical applications, CdSe foroptical applications, and borates for boron neutron capture treatment.

In some embodiments, the liquid material is selected from one of aresist polymer and a low-k dielectric. In some embodiments, the liquidmaterial comprises a non-wetting agent.

In some embodiments, the disposing of the volume of liquid material isregulated by a spreading process. In some embodiments, the spreadingprocess comprises:

-   -   (a) disposing a first volume of liquid material on the patterned        template to form a layer of liquid material on the patterned        template; and    -   (b) drawing an implement across the layer of liquid material to:        -   (i) remove a second volume of liquid material from the layer            of liquid material on the patterned template; and        -   (ii) leave a third volume of liquid material on the            patterned template.            In some embodiments, the contacting of the first template            surface with the substrate eliminates essentially all of the            disposed volume of liquid material.

In some embodiments, the treating of the liquid material comprises aprocess selected from the group consisting of a thermal process, aphotochemical process, and a chemical process.

In some embodiments, the method comprises a batch process. In someembodiments, the batch process is selected from one of a semi-batchprocess and a continuous batch process.

In some embodiments, the presently disclosed subject matter describes apatterned substrate formed by the presently disclosed methods.

VIII. Imprint Lithography Free of a Residual “Scum Layer”

A characteristic of imprint lithography that has restrained its fullpotential is the formation of a “scum layer” once the liquid material,e.g., a resin, is patterned. The “scum layer” comprises residual liquidmaterial that remains between the stamp and the substrate. In someembodiments, the presently disclosed subject matter provides a processfor generating patterns essentially free of a scum layer.

Referring now to FIGS. 9A-9E, in some embodiments, a method for forminga pattern on a substrate is provided, wherein the pattern is essentiallyfree of a scum layer. Referring now to FIG. 9A, a patterned template 910is provided. Patterned template 910 further comprises a first patternedtemplate surface 912 and a second template surface 914. The firstpatterned template surface 912 further comprises a plurality of recesses916. In some embodiments, a non-wetting agent 960 is disposed on thefirst patterned template surface 912.

Referring again to FIG. 9A, a substrate 920 is provided. Substrate 920comprises a substrate surface 922. In some embodiments, a non-wettingagent 960 is disposed on substrate surface 920.

In some embodiments, as illustrated in FIG. 9A, patterned template 910and substrate 920 are positioned in a spaced relationship to each othersuch that first patterned template surface 912 faces substrate surface922 and a gap 930 is created between first patterned template surface912 and substrate surface 922.

Referring now to FIG. 9B, a volume of liquid material 940 is disposed inthe gap 930 between first patterned template surface 912 and substratesurface 922. In some embodiments, the volume of liquid material 940 isdisposed directly on first patterned template surface 912. In someembodiments, the volume of liquid material 940 is disposed directly onnon-wetting agent 960, which is disposed on first patterned templatesurface 912. In some embodiments, the volume of liquid material 940 isdisposed directly on substrate surface 920. In some embodiments, thevolume of liquid material 940 is disposed directly on non-wetting agent960, which is disposed on substrate surface 920.

Referring now to FIG. 9C, in some embodiments, first patterned templatesurface 912 is contacted with the volume of liquid material 940. A forceF_(a) is applied to second template surface 914 thereby forcing thevolume of liquid material 940 into the plurality of recesses 916. Incontrast with the embodiment illustrated in FIG. 9 , a portion of thevolume of liquid material 940 is forced out of gap 930 by force F_(o)when force F_(a) is applied.

Referring again to FIG. 9C, in some embodiments, the volume of liquidmaterial 940 is treated by a treating process T_(r) while force F_(a) isbeing applied to form a treated liquid material 942.

Referring now to FIG. 9D, a force F_(r) is applied to patterned template910 to remove patterned template 910 from treated liquid material 942 toreveal a pattern 950 on substrate 920 as shown in FIG. 9E. In thisembodiment, substrate 920 is essentially free of a residual, or “scum,”layer of treated liquid material 942.

In some embodiments, at least one of the template surface and substratecomprises a functionalized surface element. In some embodiments, thefunctionalized surface element is functionalized with a non-wettingmaterial. In some embodiments, the non-wetting material comprisesfunctional groups that bind to the liquid material. In some embodiments,the non-wetting material is selected from the group consisting of atrichloro silane, a trialkoxy silane, a trichloro silane comprisingnon-wetting and reactive functional groups, a trialkoxy silanecomprising non-wetting and reactive functional groups, and mixturesthereof.

In some embodiments, the point of contact between the two surfaceelements is free of liquid material. In some embodiments, the point ofcontact between the two surface elements comprises residual liquidmaterial. In some embodiments, the height of the residual liquidmaterial is less than 30% of the height of the structure. In someembodiments, the height of the residual liquid material is less than 20%of the height of the structure. In some embodiments, the height of theresidual liquid material is less than 10% of the height of thestructure. In some embodiments, the height of the residual liquidmaterial is less than 5% of the height of the structure. In someembodiments, the volume of liquid material is less than the volume ofthe patterned template. In some embodiments, substantially all of thevolume of liquid material is confined to the patterned template of atleast one of the surface elements. In some embodiments, having the pointof contact between the two surface elements free of liquid materialretards slippage between the two surface elements.

IX. Solvent-Assisted Micro-Molding (SAMIM)

In some embodiments, the presently disclosed subject matter describes asolvent-assisted micro-molding (SAMIM) method for forming a pattern on asubstrate.

Referring now to FIG. 10A, a patterned template 1010 is provided.Patterned template 1010 further comprises a first patterned templatesurface 1012 and a second template surface 1014. The first patternedtemplate surface 1012 further comprises a plurality of recesses 1016.

Referring again to FIG. 10A, a substrate 1020 is provided. Substrate1020 comprises a substrate surface 1022. In some embodiments, apolymeric material 1070 is disposed on substrate surface 1022. In someembodiments, polymeric material 1070 comprises a resist polymer.

Referring again to FIG. 10A, patterned template 1010 and substrate 1020are positioned in a spaced relationship to each other such that firstpatterned template surface 1012 faces substrate surface 1022 and a gap1030 is created between first patterned template surface 1012 andsubstrate surface 1022. As shown in FIG. 10A, a solvent S is disposedwithin gap 1030, such that solvent S contacts polymeric material 1070forming a swollen polymeric material 1072.

Referring now to FIGS. 10B and 10C, first patterned template surface1012 is contacted with swollen polymeric material 1072. A force F_(a) isapplied to second template surface 1014 thereby forcing a portion ofswollen polymeric material 1072 into the plurality of recesses 1016 andleaving a portion of swollen polymeric material 1072 between firstpatterned template surface 1012 and substrate surface 1020. The swollenpolymeric material 1072 is then treated by a treating process T_(r)while under pressure.

Referring now to FIG. 10D, a force F_(r) is applied to patternedtemplate 1010 to remove patterned template 1010 from treated swollenpolymeric material 1072 to reveal a polymeric pattern 1074 on substrate1020 as shown in FIG. 10E.

X. Removing the Patterned Structure from the Patterned Template and/orSubstrate

In some embodiments, the patterned structure (e.g., a patterned micro-or nanostructure) is removed from at least one of the patterned templateand/or the substrate. This can be accomplished by a number ofapproaches, including but not limited to applying the surface elementcontaining the patterned structure to a surface that has an affinity forthe patterned structure; deforming the surface element containing thepatterned structure such that the patterned structure is released fromthe surface element; swelling the surface element containing thepatterned structure with a first solvent to extrude the patternedstructure; and washing the surface element containing the patternedstructure with a second solvent that has an affinity for the patternedstructure.

In some embodiments, the first solvent comprises supercritical fluidcarbon dioxide. In some embodiments, the first solvent comprises water.In some embodiments, the first solvent comprises an aqueous solutioncomprising water and a detergent. In embodiments, the deforming thesurface element is performed by applying a mechanical force to thesurface element. In some embodiments, the method of removing thepatterned structure further comprises a sonication method.

XI. Method of Fabricating Molecules and for Delivering a TherapeuticAgent to a Target

In some embodiments, the presently disclosed subject matter describesmethods and processes, and products by processes, for fabricating“molecules,” for use in drug discovery and drug therapies. In someembodiments, the method or process for fabricating a molecule comprisesa combinatorial method or process. In some embodiments, the method forfabricating molecules comprises a non-wetting imprint lithographymethod.

XI.A Method of Fabricating Molecules

In some embodiments, the non-wetting imprint lithography method furthercomprises a surface derived from or comprising a solvent resistant, lowsurface energy polymeric material derived from casting low viscosityliquid materials onto a master template and then curing the lowviscosity liquid materials to generate a patterned template. In someembodiments, the surface comprises a solvent resistant elastomericmaterial.

In some embodiments, the non-wetting imprint lithography method is usedto generate isolated structures. In some embodiments, the isolatedstructures comprise isolated micro-structures. In some embodiments, theisolated structures comprise isolated nano-structures. In someembodiments, the isolated structures comprise a biodegradable material.In some embodiments, the isolated structures comprise a hydrophilicmaterial. In some embodiments, the isolated structures comprise ahydrophobic material. In some embodiments, the isolated structurescomprise a particular shape. In some embodiments, the isolatedstructures further comprise “cargo.”

In some embodiments, the non-wetting imprint lithography method furthercomprises adding molecular modules, fragments, or domains to thesolution to be molded. In some embodiments, the molecular modules,fragments, or domains impart functionality to the isolated structures.In some embodiments, the functionality imparted to the isolatedstructure comprises a therapeutic functionality.

In some embodiments, a therapeutic agent, such as a drug, isincorporated into the isolated structure. In some embodiments, thephysiologically active drug is tethered to a linker to facilitate itsincorporation into the isolated structure. In some embodiments, thedomain of an enzyme or a catalyst is added to the isolated structure. Insome embodiments, a ligand or an oligopeptide is added to the isolatedstructure. In some embodiments, the oligopeptide is functional. In someembodiments, the functional oligopeptide comprises a cell targetingpeptide. In some embodiments, the functional oligopeptide comprises acell penetrating peptide. In some embodiments an antibody or functionalfragment thereof is added to the isolated structure.

In some embodiments, a binder is added to the isolated structure. Insome embodiments, the isolated structure comprising the binder is usedto fabricate identical structures. In some embodiments, the isolatedstructure comprising the binder is used to fabricate structures of avarying structure. In some embodiments, the structures of a varyingstructure are used to explore the efficacy of a molecule as atherapeutic agent. In some embodiments, the shape of the isolatedstructure mimics a biological agent. In some embodiments, the methodfurther comprises a method for drug discovery.

XIB. Method of Delivering a Therapeutic Agent to a Target

In some embodiments, a method of delivering a therapeutic agent to atarget is disclosed, the method comprising: providing a particleproduced as described herein; admixing the therapeutic agent with theparticle; and delivering the particle comprising the therapeutic agentto the target.

In some embodiments, the therapeutic agent comprises a drug. In someembodiments, the therapeutic agent comprises genetic material. In someembodiments, the genetic material is selected from the group consistingof a non-viral gene vector, DNA, RNA, RNAi, and a viral particle.

In some embodiments, the particle has a diameter of less than 100microns. In some embodiments, the particle has a diameter of less than10 microns. In some embodiments, the particle has a diameter of lessthan 1 micron. In some embodiments, the particle has a diameter of lessthan 100 nm. In some embodiments, the particle has a diameter of lessthan 10 nm.

In some embodiments, the particle comprises a biodegradable polymer. Insome embodiments, the biodegradable polymer is selected from the groupconsisting of a polyester, a polyanhydride, a polyamide, aphosphorous-based polymer, a poly(cyanoacrylate), a polyurethane, apolyorthoester, a polydihydropyran, and a polyacetal. In someembodiments, the polyester is selected from the group consisting ofpolylactic acid, polyglycolic acid, poly(hydroxybutyrate),poly(ε-caprolactone), poly(β-malic acid), and poly(dioxanones). In someembodiments, the polyanhydride is selected from the group consisting ofpoly(sebacic acid), poly(adipic acid), and poly(terpthalic acid). Insome embodiments, the polyamide is selected from the group consisting ofpoly(imino carbonates) and polyaminoacids. In some embodiments, thephosphorous-based polymer is selected from the group consisting ofpolyphosphates, polyphosphonates, and polyphosphazenes. In someembodiments, the polymer is responsive to stimuli, such as pH,radiation, ionic strength, temperature, and alternating magnetic orelectric fields.

Responses to such stimuli can include swelling and/or heating, which canfacilitate release of its cargo, or degradation.

In some embodiments, the presently disclosed subject matter describesmagneto containing particles for applications in hyperthermia therapy,cancer and gene therapy, drug delivery, magnetic resonance imagingcontrast agents, vaccine adjuvants, memory devices, and spintronics.

Without being bound to any one particular theory, the magneto containingparticles, e.g., a magnetic nanoparticle, produce heat by the process ofhyperthermia (between 41 and 46° C.) or thermo ablation (greater than46° C.), i.e., the controlled heating of the nanoparticles upon exposureto an AC-magnetic field. The heat is used to (i) induce a phase changein the polymer component (for example melt and release an encapsulatedmaterial) and/or (ii) hyperthermia treatment of specific cells and/or(iii) increase the effectiveness of the encapsulated material. Thetriggering mechanism of the magnetic nanoparticles via electromagneticheating enhance the (iv) degradation rate of the particulate; (v) caninduce swelling; and/or (vi) induce dissolution/phase change that canlead to a greater surface area, which can be beneficial when treating avariety of diseases.

In some embodiments, the presently disclosed subject matter describes analternative therapeutic agent delivery method, which utilizes“non-wetting” imprint lithography to fabricate monodisperse magneticnanoparticles for use in a drug delivery system. Such particles can beused for: (1) hyperthermia treatment of cancer cells; (2) MRI contrastagents; (3) guided delivery of the particle; and (4) triggereddegradation of the drug delivery vector.

In some embodiments, the therapeutic agent delivery system comprises abiocompatible material and a magnetic nanoparticle. In some embodiments,the biocompatible material has a melting point below 100° C. In someembodiments, the biocompatible material is selected from the groupconsisting of, but not limited to, a polylactide, a polyglycolide, ahydroxypropylcellulose, and a wax.

In some embodiments, once the magnetic nanoparticle is delivered to thetarget or is in close proximity to the target, the magnetic nanoparticleis exposed to an AC-magnetic field. The exposure to the AC-magneticfield causes the magnetic nanoparticle to undergo a controlled heating.Without being bound to any one particular theory, the controlled heatingis a result of a thermo ablation process. In some embodiments, the heatis used to induce a phase change in the polymer component of thenanoparticle. In some embodiments, the phase change comprises a meltingprocess. In some embodiments, the phase change results in the release ofan encapsulated material. In some embodiments, the release of anencapsulated material comprises a controlled release. In someembodiments, the controlled release of the encapsulated material resultsin a concentrated dosing of the therapeutic agent. In some embodiments,the heating results in the hyperthermic treatment of the target, e.g.,specific cells. In some embodiments, the heating results in an increasein the effectiveness of the encapsulated material. In some embodiments,the triggering mechanism of the magnetic nanoparticles induced by theelectromagnetic heating enhances the degradation rate of the particleand can induce swelling and/or a dissolution/phase change that can leadto a greater surface area which can be beneficial when treating avariety of diseases.

In some embodiments, additional components, including drugs, such as ananticancer agent, e.g., nitrogen mustard, cisplatin, and doxorubicin;targeting ligands, such as cell-targeting peptides, cell-penetratingpeptides, integrin receptor peptide (GRGDSP), melanocyte stimulatinghormone, vasoactive intestinal peptide, anti-Her2 mouse antibodies, anda variety of vitamins; viruses, polysaccharides, cyclodextrins,proteins, liposomes, optical nanoparticles, such as CdSe for opticalapplications, and borate nanoparticles to aid in boron neutron capturetherapy (BNCT) targets.

The presently described magnetic containing materials also lendthemselves to other applications. The magneto-particles can be assembledinto well-defined arrays driven by their shape, functionalization of thesurface and/or exposure to a magnetic field for investigations of andnot limited to magnetic assay devices, memory devices, spintronicapplications, and separations of solutions.

Thus, the presently disclosed subject matter provides a method fordelivering a therapeutic agent to a target, the method comprising:

-   -   (a) providing a particle prepared by the presently disclosed        methods;    -   (b) admixing the therapeutic agent with the particle; and    -   (c) delivering the particle comprising the therapeutic agent to        the target.

In some embodiments, the therapeutic agent is selected from one of adrug and genetic material. In some embodiments, the genetic material isselected from the group consisting of a non-viral gene vector, DNA, RNA,RNAi, and a viral particle.

In some embodiments, the particle comprises a biodegradable polymer. Insome embodiments, the biodegradable polymer is selected from the groupconsisting of a polyester, a polyanhydride, a polyamide, aphosphorous-based polymer, a poly(cyanoacrylate), a polyurethane, apolyorthoester, a polydihydropyran, and a polyacetal.

In some embodiments, the polyester is selected from the group consistingof polylactic acid, polyglycolic acid, poly(hydroxybutyrate),poly(ε-caprolactone), poly(β-malic acid), and poly(dioxanones).

In some embodiments, the polyanhydride is selected from the groupconsisting of poly(sebacic acid), poly(adipic acid), and poly(terpthalicacid).

In some embodiments, the polyamide is selected from the group consistingof poly(imino carbonates) and polyaminoacids.

In some embodiments, the phosphorous-based polymer is selected from thegroup consisting of a polyphosphate, a polyphosphonate, and apolyphosphazene.

In some embodiments, the biodegradable polymer further comprises apolymer that is responsive to a stimulus. In some embodiments, thestimulus is selected from the group consisting of pH, radiation, ionicstrength, temperature, an alternating magnetic field, and an alternatingelectric field. In some embodiments, the stimulus comprises analternating magnetic field.

In some embodiments, the method comprises exposing the particle to analternating magnetic field once the particle is delivered to the target.In some embodiments, the exposing of the particle to an alternatingmagnetic field causes the particle to produce heat through one of ahypothermia process and a thermo ablation process.

In some embodiments, the heat produced by the particle induces one of aphase change in the polymer component of the particle and a hyperthermictreatment of the target. In some embodiments, the phase change in thepolymer component of the particle comprises a change from a solid phaseto a liquid phase. In some embodiments, the phase change from a solidphase to a liquid phase causes the therapeutic agent to be released fromthe particle. In some embodiments, the release of the therapeutic agentfrom the particle comprises a controlled release.

In some embodiments, the target is selected from the group consisting ofa cell-targeting peptide, a cell-penetrating peptide, an integrinreceptor peptide (GRGDSP), a melanocyte stimulating hormone, avasoactive intestinal peptide, an anti-Her2 mouse antibody, and avitamin.

With respect to the methods of the presently disclosed subject matter,any animal subject can be treated. The term “subject” as used hereinrefers to any vertebrate species. The methods of the presently claimedsubject matter are particularly useful in the diagnosis of warm-bloodedvertebrates. Thus, the presently claimed subject matter concernsmammals. In some embodiments provided is the diagnosis and/or treatmentof mammals such as humans, as well as those mammals of importance due tobeing endangered (such as Siberian tigers), of economical importance(animals raised on farms for consumption by humans) and/or socialimportance (animals kept as pets or in zoos) to humans, for instance,carnivores other than humans (such as cats and dogs), swine (pigs, hogs,and wild boars), ruminants (such as cattle, oxen, sheep, giraffes, deer,goats, bison, and camels), and horses. Also provided is the diagnosisand/or treatment of livestock, including, but not limited todomesticated swine (pigs and hogs), ruminants, horses, poultry, and thelike.

The following references are incorporated herein by reference in theirentirety. Published International PCT Application No. WO2004081666 toDeSimone et al.; U.S. Pat. No. 6,528,080 to Dunn et al.; U.S. Pat. No.6,592,579 to Arndt et al., Published International PCT Application No.WO0066192 to Jordan; Hilger, I. et al., Radiology 570-575 (2001);Mornet, S. et al, J. Mat. Chem., 2161-2175 (2004); Berry, C. C. et al.,J. Phys. D: Applied Physics 36, R198-R206 (2003); Babincova, M. et al.,Bioelectrochemistry 55, 17-19 (2002); Wolf, S. A. et al., Science 16,1488-1495 (2001); and Sun, S. et al., Science 287, 1989-1992 (2000);U.S. Pat. No. 6,159,443 to Hallahan; and Published PCT Application No.WO 03/066066 to Hallahan et al.

XII. Method of Patterning Natural and Synthetic Structures

In some embodiments, the presently disclosed subject matter describesmethods and processes, and products by processes, for generatingsurfaces and molds from natural structures, single molecules, orself-assembled structures. Accordingly, in some embodiments, thepresently disclosed subject matter describes a method of patterning anatural structure, single molecule, and/or a self-assembled structure.In some embodiments, the method further comprises replicating thenatural structure, single molecule, and/or a self-assembled structure.In some embodiments, the method further comprises replicating thefunctionality of the natural structure, single molecule, and/or aself-assembled structure.

More particularly, in some embodiments, the method further comprisestaking the impression or mold of a natural structure, single molecule,and/or a self-assembled structure. In some embodiments, the impressionor mold is taken with a low surface energy polymeric precursor. In someembodiments, the low surface energy polymeric precursor comprises aperfluoropolyether (PFPE) functionally terminated diacrylate. In someembodiments, the natural structure, single molecule, and/orself-assembled structure is selected from the group consisting ofenzymes, viruses, antibodies, micelles, and tissue surfaces.

In some embodiments, the impression or mold is used to replicate thefeatures of the natural structure, single molecule, and/or aself-assembled structure into an isolated object or a surface. In someembodiments, a non-wetting imprint lithography method is used to impartthe features into a molded part or surface. In some embodiments, themolded part or surface produced by this process can be used in manyapplications, including, but not limited to, drug delivery, medicaldevices, coatings, catalysts, or mimics of the natural structures fromwhich they are derived. In some embodiments, the natural structurecomprises biological tissue. In some embodiments, the biological tissuecomprises tissue from a bodily organ, such as a heart. In someembodiments, the biological tissue comprises vessels and bone. In someembodiments, the biological tissue comprises tendon or cartilage. Forexample, in some embodiments, the presently disclosed subject matter canbe used to pattern surfaces for tendon and cartilage repair. Such repairtypically requires the use of collagen tissue, which comes from cadaversand must be machined for use as replacements. Most of these replacementsfail because one cannot lay down the primary pattern that is requiredfor replacement. The soft lithographic methods described hereinalleviate this problem.

In some embodiments, the presently disclosed subject matter can beapplied to tissue regeneration using stem cells. Almost all stem cellapproaches known in the art require molecular patterns for the cells toseed and then grow, thereby taking the shape of an organ, such as aliver, a kidney, or the like. In some embodiments, the molecularscaffold is cast and used as crystals to seed an organ in a form oftransplant therapy. In some embodiments, the stem cell andnano-substrate is seeded into a dying tissue, e.g., liver tissue, topromote growth and tissue regeneration. In some embodiments, thematerial to be replicated in the mold comprises a material that issimilar to or the same as the material that was originally molded. Insome embodiments, the material to be replicated in the mold comprises amaterial that is different from and/or has different properties than thematerial that was originally molded. This approach could play animportant role in addressing the organ transplant shortage.

In some embodiments, the presently disclosed subject matter is used totake the impression of one of an enzyme, a bacterium, and a virus. Insome embodiments, the enzyme, bacterium, or virus is then replicatedinto a discrete object or onto a surface that has the shape reminiscentof that particular enzyme, bacterium, or virus replicated into it. Insome embodiments, the mold itself is replicated on a surface, whereinthe surface-attached replicated mold acts as a receptor site for anenzyme, bacterium, or virus particle. In some embodiments, thereplicated mold is useful as a catalyst, a diagnostic sensor, atherapeutic agent, a vaccine, and the like. In some embodiments, thesurface-attached replicated mold is used to facilitate the discovery ofnew therapeutic agents.

In some embodiments, the macromolecular, e.g., enzyme, bacterial, orviral, molded “mimics” serve as non-self-replicating entities that havethe same surface topography as the original macromolecule, bacterium, orvirus. In some embodiments, the molded mimics are used to createbiological responses, e.g., an allergic response, to their presence,thereby creating antibodies or activating receptors. In someembodiments, the molded mimics function as a vaccine. In someembodiments, the efficacy of the biologically-active shape of the moldedmimics is enhanced by a surface modification technique.

XIII. Method of Modifying the Surface of an Imprint Lithography Mold toImpart Surface Characteristics to Molded Products

In some embodiments, the presently disclosed subject matter describes amethod of modifying the surface of an imprint lithography mold. In someembodiments, the method further comprises imparting surfacecharacteristics to a molded product. In some embodiments, the moldedproduct comprises an isolated molded product. In some embodiments, theisolate molded product is formed using a non-wetting imprint lithographytechnique. In some embodiments, the molded product comprises a contactlens, a medical device, and the like.

More particularly, the surface of a solvent resistant, low surfaceenergy polymeric material, or more particularly a PFPE mold is modifiedby a surface modification step, wherein the surface modification step isselected from the group consisting of plasma treatment, chemicaltreatment, and the adsorption of molecules. In some embodiments, themolecules adsorbed during the surface modification step are selectedfrom the group consisting of polyelectrolytes, poly(vinylalcohol),alkylhalosilanes, and ligands. In some embodiments, the structures,particles, or objects obtained from the surface-treated molds can bemodified by the surface treatments in the mold. In some embodiments, themodification comprises the pre-orientation of molecules or moieties withthe molecules comprising the molded products. In some embodiments, thepre-orientation of the molecules or moieties imparts certain propertiesto the molded products, including catalytic, wettable, adhesive,non-stick, interactive, or not interactive, when the molded product isplaced in another environment. In some embodiments, such properties areused to facilitate interactions with biological tissue or to preventinteraction with biological tissues. Applications of the presentlydisclosed subject matter include sensors, arrays, medical implants,medical diagnostics, disease detection, and separation media.

XIV. Methods for Selectively Exposing the Surface of an Article to anAgent

Also disclosed herein is a method for selectively exposing the surfaceof an article to an agent. In some embodiments the method comprises:

-   -   (a) shielding a first portion of the surface of the article with        a masking system, wherein the masking system comprises a        elastomeric mask in conformal contact with the surface of the        article; and    -   (b) applying an agent to be patterned within the masking system        to a second portion of the surface of the article, while        preventing application of the agent to the first portion        shielded by the masking system.

In some embodiments, the elastomeric mask comprises a plurality ofchannels. In some embodiments, each of the channels has across-sectional dimension of less than about 1 millimeter. In someembodiments, each of the channels has a cross-sectional dimension ofless than about 1 micron. In some embodiments, each of the channels hasa cross-sectional dimension of less than about 100 nm. In someembodiments, each of the channels has a cross-sectional dimension ofabout 1 nm. In some embodiments, the agent swells the elastomeric maskless than 25%.

In some embodiments, the agent comprises an organic electroluminescentmaterial or a precursor thereof. In some embodiments, the method furthercomprising allowing the organic electroluminescent material to form fromthe agent at the second portion of the surface, and establishingelectrical communication between the organic electroluminescent materialand an electrical circuit.

In some embodiments, the agent comprises a liquid or is carried in aliquid. In some embodiments, the agent comprises the product of chemicalvapor deposition. In some embodiments, the agent comprises a product ofdeposition from a gas phase. In some embodiments, the agent comprises aproduct of e-beam deposition, evaporation, or sputtering. In someembodiments, the agent comprises a product of electrochemicaldeposition. In some embodiments, the agent comprises a product ofelectroless deposition. In some embodiments, the agent is applied from afluid precursor. In some embodiments, comprises a solution or suspensionof an inorganic compound. In some embodiments, the inorganic compoundhardens on the second portion of the article surface.

In some embodiments, the fluid precursor comprises a suspension ofparticles in a fluid carrier. In some embodiments, the method furthercomprises allowing the fluid carrier to dissipate thereby depositing theparticles at the first region of the article surface. In someembodiments, the fluid precursor comprises a chemically active agent ina fluid carrier. In some embodiments, the method further comprisesallowing the fluid carrier to dissipate thereby depositing thechemically active agent at the first region of the article surface.

In some embodiments, the chemically active agent comprises a polymerprecursor. In some embodiments, the method further comprises forming apolymeric article from the polymer precursor. In some embodiments, thechemically active agent comprises an agent capable of promotingdeposition of a material. In some embodiments, the chemically activeagent comprises an etchant. In some embodiments, the method furthercomprises allowing the second portion of the surface of the article tobe etched. In some embodiments, the method further comprises removingthe elastomeric mask of the masking system from the first portion of thearticle surface while leaving the agent adhered to the second portion ofthe article surface.

XV. Methods for Forming Engineered Membranes

The presently disclosed subject matter also describes a method forforming an engineered membrane. In some embodiments, a patternednon-wetting template is formed by contacting a first liquid material,such as a PFPE material, with a patterned substrate and treating thefirst liquid material, for example, by curing through exposure to UVlight to form a patterned non-wetting template. The patterned substratecomprises a plurality of recesses or cavities configured in a specificshape such that the patterned non-wetting template comprises a pluralityof extruding features. The patterned non-wetting template is contactedwith a second liquid material, for example, a photocurable resin. Aforce is then applied to the patterned non-wetting template to displacean excess amount of second liquid material or “scum layer.” The secondliquid material is then treated, for example, by curing through exposureto UV light to form an interconnected structure comprising a pluralityof shape and size specific holes. The interconnected structure is thenremoved from the non-wetting template. In some embodiments, theinterconnected structure is used as a membrane for separations.

XVI. Methods for Inspecting Processes and Products by Processes

It will be important to inspect the objects/structures/particlesdescribed herein for accuracy of shape, placement and utility. Suchinspection can allow for corrective actions to be taken or for defectsto be removed or mitigated. The range of approaches and monitoringdevices useful for such inspections include: air gages, which usepneumatic pressure and flow to measure or sort dimensional attributes;balancing machines and systems, which dynamically measure and/or correctmachine or component balance; biological microscopes, which typicallyare used to study organisms and their vital processes; bore and IDgages, which are designed for internal diameter dimensional measurementor assessment; boroscopes, which are inspection tools with rigid orflexible optical tubes for interior inspection of holes, bores,cavities, and the like; calipers, which typically use a precise slidemovement for inside, outside, depth or step measurements, some of whichare used for comparing or transferring dimensions; CMM probes, which aretransducers that convert physical measurements into electrical signals,using various measuring systems within the probe structure; color andappearance instruments, which, for example, typically are used tomeasure the properties of paints and coatings including color, gloss,haze and transparency; color sensors, which register items by contrast,true color, or translucent index, and are based on one of the colormodels, most commonly the RGB model (red, green, blue); coordinatemeasuring machines, which are mechanical systems designed to move ameasuring probe to determine the coordinates of points on a work piecesurface; depth gages, which are used to measure of the depth of holes,cavities or other component features; digital/video microscopes, whichuse digital technology to display the magnified image; digital readouts,which are specialized displays for position and dimension readings frominspection gages and linear scales, or rotary encoders on machine tools;dimensional gages and instruments, which provide quantitativemeasurements of a product's or component's dimensional and formattributes such as wall thickness, depth, height, length, I.D., O.D.,taper or bore; dimensional and profile scanners, which gathertwo-dimensional or three-dimensional information about an object and areavailable in a wide variety of configurations and technologies; electronmicroscopes, which use a focused beam of electrons instead of light to“image” the specimen and gain information as to its structure andcomposition; fiberscopes, which are inspection tools with flexibleoptical tubes for interior inspection of holes, bores, and cavities;fixed gages, which are designed to access a specific attribute based oncomparative gaging, and include Angle Gages, Ball Gages, Center Gages,Drill Size Gages, Feeler Gages, Fillet Gages, Gear Tooth Gages, Gage orShim Stock, Pipe Gages, Radius Gages, Screw or Thread Pitch Gages, TaperGages, Tube Gages, U.S. Standard Gages (Sheet/Plate), Weld Gages andWire Gages; specialty/form gages, which are used to inspect parameterssuch as roundness, angularity, squareness, straightness, flatness,runout, taper and concentricity; gage blocks, which are manufactured toprecise gagemaker tolerance grades for calibrating, checking, andsetting fixed and comparative gages; height gages, which are used formeasuring the height of components or product features; indicators andcomparators, which measure where the linear movement of a precisionspindle or probe is amplified; inspection and gaging accessories, suchas layout and marking tolls, including hand tools, supplies andaccessories for dimensional measurement, marking, layout or othermachine shop applications such as scribes, transfer punches, dividers,and layout fluid; interferometers, which are used to measure distance interms of wavelength and to determine wavelengths of particular lightsources; laser micrometers, which measure extremely small distancesusing laser technology; levels, which are mechanical or electronic toolsthat measure the inclination of a surface relative to the earth'ssurface; machine alignment equipment, which is used to align rotating ormoving parts and machine components; magnifiers, which are inspectioninstruments that are used to magnify a product or part detail via a lenssystem; master and setting gages, which provide dimensional standardsfor calibrating other gages; measuring microscopes, which are used bytoolmakers for measuring the properties of tools, and often are used fordimensional measurement with lower magnifying powers to allow forbrighter, sharper images combined with a wide field of view;metallurgical microscopes, which are used for metallurgical inspection;micrometers, which are instruments for precision dimensional gagingconsisting of a ground spindle and anvil mounted in a C-shaped steelframe. Noncontact laser micrometers are also available; microscopes (alltypes), which are instruments that are capable of producing a magnifiedimage of a small object; optical/light microscopes, which use thevisible or near-visible portion of the electromagnetic spectrum; opticalcomparators, which are instruments that project a magnified image orprofile of a part onto a screen for comparison to a standard overlayprofile or scale; plug/pin gages, which are used for a “go/no-go”assessment of hole and slot dimensions or locations compared tospecified tolerances; protractors and angle gages, which measure theangle between two surfaces of a part or assembly; ring gages, which areused for “go/no-go” assessment compared to the specified dimensionaltolerances or attributes of pins, shafts, or threaded studs; rules andscales, which are flat, graduated scales used for length measurement,and which for OEM applications, digital or electronic linear scales areoften used; snap gages, which are used in production settings wherespecific diametrical or thickness measurements must be repeatedfrequently with precision and accuracy; specialty microscopes, which areused for specialized applications including metallurgy, gemology, or usespecialized techniques like acoustics or microwaves to perform theirfunction; squares, which are used to indicate if two surfaces of a partor assembly are perpendicular; styli, probes, and cantilevers, which areslender rod-shaped stems and contact tips or points used to probesurfaces in conjunction with profilometers, SPMs, CMMs, gages anddimensional scanners; surface profilometers, which measure surfaceprofiles, roughness, waviness and other finish parameters by scanning amechanical stylus across the sample or through noncontact methods;thread gages, which are dimensional instruments for measuring threadsize, pitch or other parameters; and videoscopes, which are inspectiontools that capture images from inside holes, bores or cavities.

EXAMPLES

The following Examples have been included to provide guidance to one ofordinary skill in the art for practicing representative embodiments ofthe presently disclosed subject matter. In light of the presentdisclosure and the general level of skill in the art, those of skill canappreciate that the following Examples are intended to be exemplary onlyand that numerous changes, modifications, and alterations can beemployed without departing from the scope of the presently disclosedsubject matter.

Example 1 Representative Procedure for Synthesis and Curing PhotocurablePerfluoropolyethers

In some embodiments, the synthesis and curing of PFPE materials of thepresently disclosed subject matter is performed by using the methoddescribed by Rolland, J. P., et al., J. Am. Chem. Soc., 2004, 126,2322-2323. Briefly, this method involves themethacrylate-functionalization of a commercially available PFPE diol(M_(n)=3800 g/mol) with isocyanatoethyl methacrylate. Subsequentphotocuring of the material is accomplished through blending with 1 wt %of 2,2-dimethoxy-2-phenylacetophenone and exposure to UV radiation(λ=365 nm).

More particularly, in a typical preparation of perfluoropolyetherdimethacrylate (PFPE DMA), poly(tetrafluoroethyleneoxide-co-difluoromethylene oxide)α,ω diol (ZDOL, average M_(n) ca. 3,800g/mol, 95%, Aldrich Chemical Company, Milwaukee, Wis., United States ofAmerica) (5.7227 g, 1.5 mmol) was added to a dry 50 mL round bottomflask and purged with argon for 15 minutes. 2-isocyanatoethylmethacrylate (EIM, 99%, Aldrich) (0.43 mL, 3.0 mmol) was then added viasyringe along with 1,1,2-trichlorotrifluoroethane (Freon 113 99%,Aldrich) (2 mL), and dibutyltin diacetate (DBTDA, 99%, Aldrich) (50 μL).The solution was immersed in an oil bath and allowed to stir at 50° C.for 24 h. The solution was then passed through a chromatographic column(alumina, Freon 113, 2×5 cm). Evaporation of the solvent yielded aclear, colorless, viscous oil, which was further purified by passagethrough a 0.22-μm polyethersulfone filter.

In a representative curing procedure for PFPE DMA, 1 wt % of2,2-dimethoxy-2-phenyl acetophenone (DMPA, 99% Aldrich), (0.05 g, 2.0mmol) was added to PFPE DMA (5 g, 1.2 mmol) along with 2 mL Freon 113until a clear solution was formed. After removal of the solvent, thecloudy viscous oil was passed through a 0.22-μm polyethersulfone filterto remove any DMPA that did not disperse into the PFPE DMA. The filteredPFPE DMA was then irradiated with a UV source (Electro-Lite Corporation,Danbury, Conn., United States of America, UV curing chamber model no.81432-ELC-500, λ=365 nm) while under a nitrogen purge for 10 min. Thisresulted in a clear, slightly yellow, rubbery material.

Example 2 Representative Fabrication of a PFPE DMA Device

In some embodiments, a PFPE DMA device, such as a stamp, was fabricatedaccording to the method described by Rolland, J. P., et al., J. Am.Chem. Soc., 2004, 126, 2322-2323. Briefly, the PFPE DMA containing aphotoinitiator, such as DMPA, was spin coated (800 rpm) to a thicknessof 20 μm onto a Si wafer containing the desired photoresist pattern.This coated wafer was then placed into the UV curing chamber andirradiated for 6 seconds. Separately, a thick layer (about 5 mm) of thematerial was produced by pouring the PFPE DMA containing photoinitiatorinto a mold surrounding the Si wafer containing the desired photoresistpattern. This wafer was irradiated with UV light for one minute.Following this, the thick layer was removed. The thick layer was thenplaced on top of the thin layer such that the patterns in the two layerswere precisely aligned, and then the entire device was irradiated for 10minutes. Once complete, the entire device was peeled from the Si waferwith both layers adhered together.

Example 3 Fabrication of Isolated Particles Using Non-Wetting ImprintLithography

3.1 Fabrication of 200-nm Trapezoidal PEG Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(See FIG. 13 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of PEG diacrylate is then placed on thetreated silicon wafer and the patterned PFPE mold placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess PEG-diacrylate. The entire apparatus isthen subjected to UV light (λ=365 nm) for ten minutes while under anitrogen purge. Particles are observed after separation of the PFPE moldand the treated silicon wafer using scanning electron microscopy (SEM)(see FIG. 14 ).

3.2 Fabrication of 500-nm Conical PEG Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 500-nm conical shapes(see FIG. 12 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of PEG diacrylate is then placed on thetreated silicon wafer and the patterned PFPE mold placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess PEG-diacrylate. The entire apparatus isthen subjected to UV light (λ=365 nm) for ten minutes while under anitrogen purge. Particles are observed after separation of the PFPE moldand the treated silicon wafer using scanning electron microscopy (SEM)(see FIG. 15 ).

3.3 Fabrication of 3-μm Arrow-Shaped PEG Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 3-μm arrow shapes (seeFIG. 11 ). A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of PEG diacrylate is then placed on thetreated silicon wafer and the patterned PFPE mold placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess PEG-diacrylate. The entire apparatus isthen subjected to UV light (λ=365 nm) for ten minutes while under anitrogen purge. Particles are observed after separation of the PFPE moldand the treated silicon wafer using scanning electron microscopy (SEM)(see FIG. 16 ).

3.4 Fabrication of 200-nm×750-nm×250-nm Rectangular PEG Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm×750-nm×250-nmrectangular shapes. A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of PEG diacrylate is then placed on thetreated silicon wafer and the patterned PFPE mold placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess PEG-diacrylate. The entire apparatus isthen subjected to UV light (λ=365 nm) for ten minutes while under anitrogen purge. Particles are observed after separation of the PFPE moldand the treated silicon wafer using scanning electron microscopy (SEM)(see FIG. 17 ).

3.5 Fabrication of 200-nm Trapezoidal Trimethylopropane Triacrylate(TMPTA) Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, TMPTA is blended with 1 wt % of a photoinitiator,1-hydroxycyclohexyl phenyl ketone. Flat, uniform, non-wetting surfacesare generated by treating a silicon wafer cleaned with “piranha”solution (1:1 concentrated sulfuric acid: 30% hydrogen peroxide (aq)solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl) silane via vapordeposition in a desiccator for 20 minutes. Following this, 50 μL ofTMPTA is then placed on the treated silicon wafer and the patterned PFPEmold placed on top of it. The substrate is then placed in a moldingapparatus and a small pressure is applied to push out excess TMPTA. Theentire apparatus is then subjected to UV light (λ=365 nm) for tenminutes while under a nitrogen purge. Particles are observed afterseparation of the PFPE mold and the treated silicon wafer using scanningelectron microscopy (SEM) (see FIG. 18 ).

3.6 Fabrication of 500-nm Conical Trimethylopropane Triacrylate (TMPTA)Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 500-nm conical shapes(see FIG. 12 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, TMPTA is blended with 1 wt % of a photoinitiator,1-hydroxycyclohexyl phenyl ketone. Flat, uniform, non-wetting surfacesare generated by treating a silicon wafer cleaned with “piranha”solution (1:1 concentrated sulfuric acid: 30% hydrogen peroxide (aq)solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl) silane via vapordeposition in a desiccator for 20 minutes. Following this, 50 μL ofTMPTA is then placed on the treated silicon wafer and the patterned PFPEmold placed on top of it. The substrate is then placed in a moldingapparatus and a small pressure is applied to push out excess TMPTA. Theentire apparatus is then subjected to UV light (λ=365 nm) for tenminutes while under a nitrogen purge. Particles are observed afterseparation of the PFPE mold and the treated silicon wafer using scanningelectron microscopy (SEM) (see FIG. 19 ). Further, FIG. 20 shows ascanning electron micrograph of 500-nm isolated conical particles ofTMPTA, which have been printed using an embodiment of the presentlydescribed non-wetting imprint lithography method and harvestedmechanically using a doctor blade. The ability to harvest particles insuch a way offers conclusive evidence for the absence of a “scum layer.”

3.7 Fabrication of 3-μm Arrow-Shaped TMPTA Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 3-μm arrow shapes (seeFIG. 11 ). A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, TMPTA is blended with 1 wt % of a photoinitiator,1-hydroxycyclohexyl phenyl ketone. Flat, uniform, non-wetting surfacesare generated by treating a silicon wafer cleaned with “piranha”solution (1:1 concentrated sulfuric acid: 30% hydrogen peroxide (aq)solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl) silane via vapordeposition in a desiccator for 20 minutes. Following this, 50 μL ofTMPTA is then placed on the treated silicon wafer and the patterned PFPEmold placed on top of it. The substrate is then placed in a moldingapparatus and a small pressure is applied to push out excess TMPTA. Theentire apparatus is then subjected to UV light (λ=365 nm) for tenminutes while under a nitrogen purge. Particles are observed afterseparation of the PFPE mold and the treated silicon wafer using scanningelectron microscopy (SEM).

3.8 Fabrication of 200-nm Trapezoidal Poly(Lactic Acid) (PLA) Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, one gram of (3S)-cis-3,6-dimethyl-1,4-dioxane-2,5-dione (LA)is heated above its melting temperature (92° C.) to 110° C. andapproximately 20 μL of stannous octoate catalyst/initiator is added tothe liquid monomer. Flat, uniform, non-wetting surfaces are generated bytreating a silicon wafer cleaned with “piranha” solution (1:1concentrated sulfuric acid: 30% hydrogen peroxide (aq) solution) withtrichloro(1H, 1H, 2H, 2H-perfluorooctyl) silane via vapor deposition ina desiccator for 20 minutes. Following this, 50 μL of molten LAcontaining catalyst is then placed on the treated silicon waferpreheated to 110° C. and the patterned PFPE mold is placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess monomer. The entire apparatus is thenplaced in an oven at 110° C. for 15 hours. Particles are observed aftercooling to room temperature and separation of the PFPE mold and thetreated silicon wafer using scanning electron microscopy (SEM) (see FIG.21 ). Further, FIG. 22 is a scanning electron micrograph of 200-nmisolated trapezoidal particles of poly(lactic acid) (PLA), which havebeen printed using an embodiment of the presently described non-wettingimprint lithography method and harvested mechanically using a doctorblade. The ability to harvest particles in such a way offers conclusiveevidence for the absence of a “scum layer.”

3.9 Fabrication of 3-μm Arrow-Shaped (PLA) Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 3-μm arrow shapes (seeFIG. 11 ). A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, one gram of (3S)-cis-3,6-dimethyl-1,4-dioxane-2,5-dione (LA)is heated above its melting temperature (92° C.) to 110° C. and ˜20 μLof stannous octoate catalyst/initiator is added to the liquid monomer.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of molten LA containing catalyst is thenplaced on the treated silicon wafer preheated to 110° C. and thepatterned PFPE mold is placed on top of it. The substrate is then placedin a molding apparatus and a small pressure is applied to push outexcess monomer. The entire apparatus is then placed in an oven at 110°C. for 15 hours. Particles are observed after cooling to roomtemperature and separation of the PFPE mold and the treated siliconwafer using scanning electron microscopy (SEM) (see FIG. 23 ).

3.10 Fabrication of 500-nm Conical Shaped (PLA) Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 500-nm conical shapes(see FIG. 12 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, one gram of (3S)-cis-3,6-dimethyl-1,4-dioxane-2,5-dione (LA)is heated above its melting temperature (92° C.) to 110° C. and ˜20 μLof stannous octoate catalyst/initiator is added to the liquid monomer.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of molten LA containing catalyst is thenplaced on the treated silicon wafer preheated to 110° C. and thepatterned PFPE mold is placed on top of it. The substrate is then placedin a molding apparatus and a small pressure is applied to push outexcess monomer. The entire apparatus is then placed in an oven at 110°C. for 15 hours. Particles are observed after cooling to roomtemperature and separation of the PFPE mold and the treated siliconwafer using scanning electron microscopy (SEM) (see FIG. 24 ).

3.11 Fabrication of 200-nm Trapezoidal Poly(Pyrrole) (Ppy) Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Separately, 50 μL of a 1:1 v:v solution oftetrahydrofuran:pyrrole is added to 50 μL of 70% perchloric acid (aq). Aclear, homogenous, brown solution quickly forms and develops into black,solid, polypyrrole in 15 minutes. A drop of this clear, brown solution(prior to complete polymerization) is placed onto a treated siliconwafer and into a stamping apparatus and a pressure is applied to removeexcess solution. The apparatus is then placed into a vacuum oven for 15h to remove the THF and water. Particles are observed using scanningelectron microscopy (SEM) (see FIG. 25 ) after release of the vacuum andseparation of the PFPE mold and the treated silicon wafer.

3.12 Fabrication of 3-μm Arrow-Shaped (Ppy) Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 3-μm arrow shapes (seeFIG. 11 ). A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master. Flat,uniform, non-wetting surfaces are generated by treating a silicon wafercleaned with “piranha” solution (1:1 concentrated sulfuric acid: 30%hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Separately, 50 μL of a 1:1 v:v solution oftetrahydrofuran:pyrrole is added to 50 μL of 70% perchloric acid (aq). Aclear, homogenous, brown solution quickly forms and develops into black,solid, polypyrrole in 15 minutes. A drop of this clear, brown solution(prior to complete polymerization) is placed onto a treated siliconwafer and into a stamping apparatus and a pressure is applied to removeexcess solution. The apparatus is then placed into a vacuum oven for 15h to remove the THF and water. Particles are observed using scanningelectron microscopy (SEM) (see FIG. 26 ) after release of the vacuum andseparation of the PFPE mold and the treated silicon wafer.

3.13 Fabrication of 500-nm Conical (Ppy) Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 500-nm conical shapes(see FIG. 12 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Separately, 50 μL of a 1:1 v:v solution oftetrahydrofuran:pyrrole is added to 50 μL of 70% perchloric acid (aq). Aclear, homogenous, brown solution quickly forms and develops into black,solid, polypyrrole in 15 minutes. A drop of this clear, brown solution(prior to complete polymerization) is placed onto a treated siliconwafer and into a stamping apparatus and a pressure is applied to removeexcess solution. The apparatus is then placed into a vacuum oven for 15h to remove the THF and water. Particles are observed using scanningelectron microscopy (SEM) (see FIG. 27 ) after release of the vacuum andseparation of the PFPE mold and the treated silicon wafer.

3.14 Encapsulation of Fluorescently Tagged DNA Inside 200-nm TrapezoidalPEG Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone. 20μL of water and 20 μL of PEG diacrylate monomer are added to 8 nanomolesof 24 bp DNA oligonucleotide that has been tagged with a fluorescentdye, CY-3. Flat, uniform, non-wetting surfaces are generated by treatinga silicon wafer cleaned with “piranha” solution (1:1 concentratedsulfuric acid: 30% hydrogen peroxide (aq) solution) with trichloro(1H,1H, 2H, 2H-perfluorooctyl) silane via vapor deposition in a desiccatorfor 20 minutes. Following this, 50 μL of the PEG diacrylate solution isthen placed on the treated silicon wafer and the patterned PFPE moldplaced on top of it. The substrate is then placed in a molding apparatusand a small pressure is applied to push out excess PEG-diacrylatesolution. The entire apparatus is then subjected to UV light (λ=365 nm)for ten minutes while under a nitrogen purge. Particles are observedafter separation of the PFPE mold and the treated silicon wafer usingconfocal fluorescence microscopy (see FIG. 28 ). Further, FIG. 28A showsa fluorescent confocal micrograph of 200 nm trapezoidal PEGnanoparticles which contain 24-mer DNA strands that are tagged withCY-3. FIG. 28B is optical micrograph of the 200-nm isolated trapezoidalparticles of PEG diacrylate that contain fluorescently tagged DNA. FIG.28C is the overlay of the images provided in FIGS. 28A and 28B, showingthat every particle contains DNA.

3.15 Encapsulation of Magnetite Nanoparticles Inside 500-nm Conical PEGParticles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 500-nm conical shapes(see FIG. 12 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Separately, citrate capped magnetite nanoparticles weresynthesized by reaction of ferric chloride (40 mL of a 1 M aqueoussolution) and ferrous chloride (10 mL of a 2 M aqueous hydrochloric acidsolution) which is added to ammonia (500 mL of a 0.7 M aqueoussolution). The resulting precipitate is collected by centrifugation andthen stirred in 2 M perchloric acid. The final solids are collected bycentrifugation. 0.290 g of these perchlorate-stabilized nanoparticlesare suspended in 50 mL of water and heated to 90° C. while stirring.Next, 0.106 g of sodium citrate is added. The solution is stirred at 90°C. for 30 min to yield an aqueous solution of citrate-stabilized ironoxide nanoparticles. 50 μL of this solution is added to 50 μL of a PEGdiacrylate solution in a microtube. This microtube is vortexed for tenseconds. Following this, 50 μL of this PEG diacrylate/particle solutionis then placed on the treated silicon wafer and the patterned PFPE moldplaced on top of it. The substrate is then placed in a molding apparatusand a small pressure is applied to push out excessPEG-diacrylate/particle solution. The entire apparatus is then subjectedto UV light (λ=365 nm) for ten minutes while under a nitrogen purge.Nanoparticle-containing PEG-diacrylate particles are observed afterseparation of the PFPE mold and the treated silicon wafer using opticalmicroscopy.

3.16 Fabrication of Isolated Particles on Glass Surfaces Using “DoubleStamping”

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone. Aflat, non-wetting surface is generated by photocuring a film of PFPE-DMAonto a glass slide, according to the procedure outlined for generating apatterned PFPE-DMA mold. 5 μL of the PEG-diacrylate/photoinitiatorsolution is pressed between the PFPE-DMA mold and the flat PFPE-DMAsurface, and pressure is applied to squeeze out excess PEG-diacrylatemonomer. The PFPE-DMA mold is then removed from the flat PFPE-DMAsurface and pressed against a clean glass microscope slide andphotocured using UV radiation (λ=365 nm) for 10 minutes while under anitrogen purge. Particles are observed after cooling to room temperatureand separation of the PFPE mold and the glass microscope slide, usingscanning electron microscopy (SEM) (see FIG. 29 ).

Example 3.17. Encapsulation of Viruses in PEG-Diacrylate Nanoparticles

A patterned perfluoropolyether (PFPE) mold is generated by pouringPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Fluorescently-labeled or unlabeled Adenovirus or Adeno-Associated Virussuspensions are added to this PEG-diacrylate monomer solution and mixedthoroughly. Flat, uniform, non-wetting surfaces are generated bytreating a silicon wafer cleaned with “piranha” solution (1:1concentrated sulfuric acid: 30% hydrogen peroxide (aq) solution) withtrichloro(1H, 1H, 2H, 2H-perfluorooctyl) silane via vapor deposition ina desiccator for 20 minutes. Following this, 50 μL of the PEGdiacrylate/virus solution is then placed on the treated silicon waferand the patterned PFPE mold placed on top of it. The substrate is thenplaced in a molding apparatus and a small pressure is applied to pushout excess PEG-diacrylate solution. The entire apparatus is thensubjected to UV light (λ=365 nm) for ten minutes while under a nitrogenpurge. Virus-containing particles are observed after separation of thePFPE mold and the treated silicon wafer using transmission electronmicroscopy or, in the case of fluorescently-labeled viruses, confocalfluorescence microscopy.

Example 3.18. Encapsulation of Proteins in PEG-Diacrylate Nanoparticles

A patterned perfluoropolyether (PFPE) mold is generated by pouringPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Fluorescently-labeled or unlabeled protein solutions are added to thisPEG-diacrylate monomer solution and mixed thoroughly. Flat, uniform,non-wetting surfaces are generated by treating a silicon wafer cleanedwith “piranha” solution (1:1 concentrated sulfuric acid: 30% hydrogenperoxide (aq) solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl)silane via vapor deposition in a desiccator for 20 minutes. Followingthis, 50 μL of the PEG diacrylate/virus solution is then placed on thetreated silicon wafer and the patterned PFPE mold placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess PEG-diacrylate solution. The entireapparatus is then subjected to UV light (λ=365 nm) for ten minutes whileunder a nitrogen purge. Protein-containing particles are observed afterseparation of the PFPE mold and the treated silicon wafer usingtraditional assay methods or, in the case of fluorescently-labeledproteins, confocal fluorescence microscopy.

Example 3.19. Fabrication of 200-nm Titania Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, 1 g of Pluronic P123 is dissolved in 12 g of absoluteethanol. This solution was added to a solution of 2.7 mL of concentratedhydrochloric acid and 3.88 mL titanium (IV) ethoxide. Flat, uniform,non-wetting surfaces are generated by treating a silicon wafer cleanedwith “piranha” solution (1:1 concentrated sulfuric acid: 30% hydrogenperoxide (aq) solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl)silane via vapor deposition in a desiccator for 20 minutes. Followingthis, 50 μL of the sol-gel solution is then placed on the treatedsilicon wafer and the patterned PFPE mold placed on top of it. Thesubstrate is then placed in a molding apparatus and a small pressure isapplied to push out excess sol-gel precursor. The entire apparatus isthen set aside until the sol-gel precursor has solidified. Particles areobserved after separation of the PFPE mold and the treated silicon waferusing scanning electron microscopy (SEM).

Example 3.20. Fabrication of 200-nm Silica Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, 2 g of Pluronic P123 is dissolved in 30 g of water and 120 gof 2 M HCl is added while stirring at 35° C. To this solution, add 8.50g of TEOS with stirring at 35° C. for 20 h. Flat, uniform, non-wettingsurfaces are generated by treating a silicon wafer cleaned with“piranha” solution (1:1 concentrated sulfuric acid: 30% hydrogenperoxide (aq) solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl)silane via vapor deposition in a desiccator for 20 minutes. Followingthis, 50 μL of the sol-gel solution is then placed on the treatedsilicon wafer and the patterned PFPE mold placed on top of it. Thesubstrate is then placed in a molding apparatus and a small pressure isapplied to push out excess sol-gel precursor. The entire apparatus isthen set aside until the sol-gel precursor has solidified. Particles areobserved after separation of the PFPE mold and the treated silicon waferusing scanning electron microscopy (SEM).

Example 3.21. Fabrication of 200-nm Europium-Doped Titania Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, 1 g of Pluronic P123 and 0.51 g of EuCl₃·6 H₂O are dissolvedin 12 g of absolute ethanol. This solution is added to a solution of 2.7mL of concentrated hydrochloric acid and 3.88 mL titanium (IV) ethoxide.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of the sol-gel solution is then placed onthe treated silicon wafer and the patterned PFPE mold placed on top ofit. The substrate is then placed in a molding apparatus and a smallpressure is applied to push out excess sol-gel precursor. The entireapparatus is then set aside until the sol-gel precursor has solidified.Particles are observed after separation of the PFPE mold and the treatedsilicon wafer using scanning electron microscopy (SEM).

Example 3.22. Encapsulation of CdSe Nanoparticles Inside 200-nm PEGParticles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13 ). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Separately, 0.5 g of sodium citrate and 2 mL of 0.04 M cadmiumperchlorate are dissolved in 45 mL of water, and the pH is adjusted toof the solution to 9 with 0.1 M NaOH. The solution is bubbled withnitrogen for 15 minutes. 2 mL of 1 M N,N-dimethylselenourea is added tothe solution and heated in a microwave oven for 60 seconds. 50 μL ofthis solution is added to 50 μL of a PEG diacrylate solution in amicrotube. This microtube is vortexed for ten seconds. 50 μL of this PEGdiacrylate/CdSe particle solution is placed on the treated silicon waferand the patterned PFPE mold placed on top of it. The substrate is thenplaced in a molding apparatus and a small pressure is applied to pushout excess PEG-diacrylate solution. The entire apparatus is thensubjected to UV light (λ=365 nm) for ten minutes while under a nitrogenpurge. PEG-diacrylate particles with encapsulated CdSe nanoparticles areobserved after separation of the PFPE mold and the treated silicon waferusing TEM or fluorescence microscopy.

Example 3.23. Synthetic Replication of Adenovirus Particles UsingNon-Wetting Imprint Lithography

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersing adenovirusparticles on a silicon wafer. This master can be used to template apatterned mold by pouring PFPE-DMA containing 1-hydroxycyclohexyl phenylketone over the patterned area of the master. A poly(dimethylsiloxane)mold is used to confine the liquid PFPE-DMA to the desired area. Theapparatus is then subjected to UV light (λ=365 nm) for 10 minutes whileunder a nitrogen purge. The fully cured PFPE-DMA mold is then releasedfrom the master. Separately, TMPTA is blended with 1 wt % of aphotoinitiator, 1-hydroxycyclohexyl phenyl ketone. Flat, uniform,non-wetting surfaces are generated by treating a silicon wafer cleanedwith “piranha” solution (1:1 concentrated sulfuric acid: 30% hydrogenperoxide (aq) solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl)silane via vapor deposition in a desiccator for 20 minutes. Followingthis, 50 μL of TMPTA is then placed on the treated silicon wafer and thepatterned PFPE mold placed on top of it. The substrate is then placed ina molding apparatus and a small pressure is applied to push out excessTMPTA. The entire apparatus is then subjected to UV light (λ=365 nm) forten minutes while under a nitrogen purge. Synthetic virus replicates areobserved after separation of the PFPE mold and the treated silicon waferusing scanning electron microscopy (SEM) or transmission electronmicroscopy (TEM).

Example 3.24. Synthetic Replication of Earthworm Hemoglobin ProteinUsing Non-Wetting Imprint Lithography

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersing earthwormhemoglobin protein on a silicon wafer. This master can be used totemplate a patterned mold by pouring PFPE-DMA containing1-hydroxycyclohexyl phenyl ketone over the patterned area of the master.A poly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the master. Separately, TMPTA is blended with1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone. Flat,uniform, non-wetting surfaces are generated by treating a silicon wafercleaned with “piranha” solution (1:1 concentrated sulfuric acid: 30%hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of TMPTA is then placed on the treatedsilicon wafer and the patterned PFPE mold placed on top of it. Thesubstrate is then placed in a molding apparatus and a small pressure isapplied to push out excess TMPTA. The entire apparatus is then subjectedto UV light (λ=365 nm) for ten minutes while under a nitrogen purge.Synthetic protein replicates are observed after separation of the PFPEmold and the treated silicon wafer using scanning electron microscopy(SEM) or transmission electron microscopy (TEM).

Example 3.25. Combinatorial Engineering of 100-nm NanoparticleTherapeutics

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 100-nm cubic shapes. Apoly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the silicon master. Separately, apoly(ethylene glycol) (PEG) diacrylate (n=9) is blended with 1 wt % of aphotoinitiator, 1-hydroxycyclohexyl phenyl ketone. Other therapeuticagents (i.e., small molecule drugs, proteins, polysaccharides, DNA,etc.), tissue targeting agents (cell penetrating peptides and ligands,hormones, antibodies, etc.), therapeutic release/transfection agents(other controlled-release monomer formulations, cationic lipids, etc.),and miscibility enhancing agents (cosolvents, charged monomers, etc.)are added to the polymer precursor solution in a combinatorial manner.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuricacid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of the combinatorially-generated particleprecursor solution is then placed on the treated silicon wafer and thepatterned PFPE mold placed on top of it. The substrate is then placed ina molding apparatus and a small pressure is applied to push out excesssolution. The entire apparatus is then subjected to UV light (λ=365 nm)for ten minutes while under a nitrogen purge. The PFPE-DMA mold is thenseparated from the treated wafer, and particles are harvested and thetherapeutic efficacy of each combinatorially generated nanoparticle isestablished. By repeating this methodology with different particleformulations, many combinations of therapeutic agents, tissue targetingagents, release agents, and other important compounds can be rapidlyscreened to determine the optimal combination for a desired therapeuticapplication.

Example 3.26 Fabrication of a Shape-Specific PEG Membrane

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 3-μm cylindrical holesthat are 5 μm deep. A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuricacid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of PEG diacrylate is then placed on thetreated silicon wafer and the patterned PFPE mold placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess PEG-diacrylate. The entire apparatus isthen subjected to UV light (λ=365 nm) for ten minutes while under anitrogen purge. An interconnected membrane is observed after separationof the PFPE mold and the treated silicon wafer using scanning electronmicroscopy (SEM). The membrane is released from the surface by soakingin water and allowing it to lift off the surface.

Example 4 Molding of Features for Semiconductor Applications

4.1 Fabrication of 140-nm Lines Separated by 70 nm in TMPTA

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 140-nm lines separated by70 nm. A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, TMPTA is blended with 1 wt % of a photoinitiator,1-hydroxycyclohexyl phenyl ketone. Flat, uniform, surfaces are generatedby treating a silicon wafer cleaned with “piranha” solution (1:1concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) andtreating the wafer with an adhesion promoter, (trimethoxysilyl propylmethacryalte). Following this, 50 μL of TMPTA is then placed on thetreated silicon wafer and the patterned PFPE mold placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to ensure a conformal contact. The entire apparatus is thensubjected to UV light (λ=365 nm) for ten minutes while under a nitrogenpurge. Features are observed after separation of the PFPE mold and thetreated silicon wafer using atomic force microscopy (AFM) (see FIG. 30).

Example 4.1. Molding of a Polystyrene Solution

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 140-nm lines separated by70 nm. A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, polystyrene is dissolved in 1 to 99 wt % of toluene. Flat,uniform, surfaces are generated by treating a silicon wafer cleaned with“piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide(aq) solution) and treating the wafer with an adhesion promoter.Following this, 50 μL of polystyrene solution is then placed on thetreated silicon wafer and the patterned PFPE mold is placed on top ofit. The substrate is then placed in a molding apparatus and a smallpressure is applied to ensure a conformal contact. The entire apparatusis then subjected to vacuum for a period of time to remove the solvent.Features are observed after separation of the PFPE mold and the treatedsilicon wafer using atomic force microscopy (AFM) and scanning electronmicroscopy (SEM).

Example 4.2. Molding of Isolated Features on Microelectronics-CompatibleSurfaces Using “Double Stamping”

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 140-nm lines separated by70 nm. A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, TMPTA is blended with 1 wt % of a photoinitiator,1-hydroxycyclohexyl phenyl ketone. A flat, non-wetting surface isgenerated by photocuring a film of PFPE-DMA onto a glass slide,according to the procedure outlined for generating a patterned PFPE-DMAmold. 50 μL of the TMPTA/photoinitiator solution is pressed between thePFPE-DMA mold and the flat PFPE-DMA surface, and pressure is applied tosqueeze out excess TMPTA monomer. The PFPE-DMA mold is then removed fromthe flat PFPE-DMA surface and pressed against a clean, flatsilicon/silicon oxide wafer and photocured using UV radiation (λ=365 nm)for 10 minutes while under a nitrogen purge. Isolated, poly(TMPTA)features are observed after separation of the PFPE mold and thesilicon/silicon oxide wafer, using scanning electron microscopy (SEM).

Example 4.3. Fabrication of 200-nm Titania Structures forMicroelectronics

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 140-nm lines separated by70 nm. A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, 1 g of Pluronic P123 is dissolved in 12 g of absoluteethanol. This solution was added to a solution of 2.7 mL of concentratedhydrochloric acid and 3.88 mL titanium (IV) ethoxide. Flat, uniform,surfaces are generated by treating a silicon/silicon oxide wafer with“piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide(aq) solution) and drying. Following this, 50 μL of the sol-gel solutionis then placed on the treated silicon wafer and the patterned PFPE moldplaced on top of it. The substrate is then placed in a molding apparatusand a small pressure is applied to push out excess sol-gel precursor.The entire apparatus is then set aside until the sol-gel precursor hassolidified. Oxide structures are observed after separation of the PFPEmold and the treated silicon wafer using scanning electron microscopy(SEM).

Example 4.4. Fabrication of 200-nm Silica Structures forMicroelectronics

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 140-nm lines separated by70 nm. A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, 2 g of Pluronic P123 is dissolved in 30 g of water and 120 gof 2 M HCl is added while stirring at 35° C. To this solution, add 8.50g of TEOS with stirring at 35° C. for 20 h. Flat, uniform, surfaces aregenerated by treating a silicon/silicon oxide wafer with “piranha”solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq)solution) and drying. Following this, 50 μL of the sol-gel solution isthen placed on the treated silicon wafer and the patterned PFPE moldplaced on top of it. The substrate is then placed in a molding apparatusand a small pressure is applied to push out excess sol-gel precursor.The entire apparatus is then set aside until the sol gel precursor hassolidified. Oxide structures are observed after separation of the PFPEmold and the treated silicon wafer using scanning electron microscopy(SEM).

Example 4.5. Fabrication of 200-nm Europium-Doped Titania Structures forMicroelectronics

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 140-nm lines separated by70 nm. A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, 1 g of Pluronic P123 and 0.51 g of EuCl₃·6 H₂O are dissolvedin 12 g of absolute ethanol. This solution was added to a solution of2.7 mL of concentrated hydrochloric acid and 3.88 mL titanium (IV)ethoxide. Flat, uniform, surfaces are generated by treating asilicon/silicon oxide wafer with “piranha” solution (1:1 concentratedsulfuric acid:30% hydrogen peroxide (aq) solution) and drying. Followingthis, 50 μL of the sol-gel solution is then placed on the treatedsilicon wafer and the patterned PFPE mold placed on top of it. Thesubstrate is then placed in a molding apparatus and a small pressure isapplied to push out excess sol-gel precursor. The entire apparatus isthen set aside until the sol-gel precursor has solidified. Oxidestructures are observed after separation of the PFPE mold and thetreated silicon wafer using scanning electron microscopy (SEM).

Example 4.6. Fabrication of Isolated “Scum Free” Features forMicroelectronics

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 140-nm lines separated by70 nm. A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, TMPTA is blended with 1 wt % of a photoinitiator,1-hydroxycyclohexyl phenyl ketone. Flat, uniform, non-wetting surfacescapable of adhering to the resist material are generated by treating asilicon wafer cleaned with “piranha” solution (1:1 concentrated sulfuricacid:30% hydrogen peroxide (aq) solution) and treating the wafer with amixture of an adhesion promoter, (trimethoxysilyl propyl methacryalte)and a non-wetting silane agent (1H, 1H, 2H, 2H-perfluorooctyltrimethoxysilane). The mixture can range from 100% of the adhesionpromoter to 100% of the non-wetting silane. Following this, 50 μL ofTMPTA is then placed on the treated silicon wafer and the patterned PFPEmold placed on top of it. The substrate is then placed in a moldingapparatus and a small pressure is applied to ensure a conformal contactand to push out excess TMPTA. The entire apparatus is then subjected toUV light (λ=365 nm) for ten minutes while under a nitrogen purge.Features are observed after separation of the PFPE mold and the treatedsilicon wafer using atomic force microscopy (AFM) and scanning electronmicroscopy (SEM).

Example 5 Molding of Natural and Engineered Templates

5.1. Fabrication of a Perfluoropolyether-Dimethacrylate (PFPE-DMA) Moldfrom a Template Generated Using Electron-Beam Lithography

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated using electron beam lithographyby spin coating a bilayer resist of 200,000 MW PMMA and 900,000 MW PMMAonto a silicon wafer with 500-nm thermal oxide, and exposing this resistlayer to an electron beam that is translating in a pre-programmedpattern. The resist is developed in 3:1 isopropanol:methyl isobutylketone solution to remove exposed regions of the resist. A correspondingmetal pattern is formed on the silicon oxide surface by evaporating 5 nmCr and 15 nm Au onto the resist covered surface and lifting off theresidual PMMA/Cr/Au film in refluxing acetone. This pattern istransferred to the underlying silicon oxide surface by reactive ionetching with CF₄/O₂ plasma and removal of the Cr/Au film in aqua regia.(FIG. 31 ). This master can be used to template a patterned mold bypouring PFPE-DMA containing 1-hydroxycyclohexyl phenyl ketone over thepatterned area of the master. A poly(dimethylsiloxane) mold is used toconfine the liquid PFPE-DMA to the desired area. The apparatus is thensubjected to UV light (λ=365 nm) for 10 minutes while under a nitrogenpurge. The fully cured PFPE-DMA mold is then released from the master.This mold can be used for the fabrication of particles using non-wettingimprint lithography as specified in Particle Fabrication Examples 3.3and 3.4.

5.2 Fabrication of a Perfluoropolyether-Dimethacrylate (PFPE-DMA) Moldfrom a Template Generated Using Photolithography.

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated using photolithography by spincoating a film of SU-8 photoresist onto a silicon wafer. This resist isbaked on a hotplate at 95° C. and exposed through a pre-patternedphotomask. The wafer is baked again at 95° C. and developed using acommercial developer solution to remove unexposed SU-8 resist. Theresulting patterned surface is fully cured at 175° C. This master can beused to template a patterned mold by pouring PFPE-DMA containing1-hydroxycyclohexyl phenyl ketone over the patterned area of the master.A poly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the master, and can be imaged by opticalmicroscopy to reveal the patterned PFPE-DMA mold (see FIG. 32 ).

5.3 Fabrication of a Perfluoropolyether-Dimethacrylate (PFPE-DMA) Moldfrom a Template Generated from Dispersed Tobacco Mosaic Virus Particles

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersing tobacco mosaicvirus (TMV) particles on a silicon wafer (FIG. 33 a ). This master canbe used to template a patterned mold by pouring PFPE-DMA containing1-hydroxycyclohexyl phenyl ketone over the patterned area of the master.A poly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the master. The morphology of the mold canthen be confirmed using Atomic Force Microscopy (FIG. 33 b ).

5.4. Fabrication of a Perfluoropolyether-Dimethacrylate (PFPE-DMA) Moldfrom a Template Generated from Block-Copolymer Micelles

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersingpolystyrene-polyisoprene block copolymer micelles on a freshly-cleavedmica surface. This master can be used to template a patterned mold bypouring PFPE-DMA containing 1-hydroxycyclohexyl phenyl ketone over thepatterned area of the master. A poly(dimethylsiloxane) mold is used toconfine the liquid PFPE-DMA to the desired area. The apparatus is thensubjected to UV light (λ=365 nm) for 10 minutes while under a nitrogenpurge. The fully cured PFPE-DMA mold is then released from the master.The morphology of the mold can then be confirmed using Atomic ForceMicroscopy (see FIG. 34 ).

5.5 Fabrication of a Perfluoropolyether-Dimethacrylate (PFPE-DMA) Moldfrom a Template Generated from Brush Polymers.

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersing poly(butylacrylate) brush polymers on a freshly-cleaved mica surface. This mastercan be used to template a patterned mold by pouring PFPE-DMA containing1-hydroxycyclohexyl phenyl ketone over the patterned area of the master.A poly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the master. The morphology of the mold canthen be confirmed using Atomic Force Microscopy (FIG. 35 ).

Example 5.6. Fabrication of a Perfluoropolyether-Dimethacrylate(PFPE-DMA) Mold from a Template Generated from Earthworm HemoglobinProtein

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersing earthwormhemoglobin proteins on a freshly-cleaved mica surface. This master canbe used to template a patterned mold by pouring PFPE-DMA containing1-hydroxycyclohexyl phenyl ketone over the patterned area of the master.A poly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the master. The morphology of the mold canthen be confirmed using Atomic Force Microscopy.

Example 5.7. Fabrication of a Perfluoropolyether-Dimethacrylate(PFPE-DMA) Mold from a Template Generated from Patterned DNANanostructures

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersing DNAnanostructures on a freshly-cleaved mica surface. This master can beused to template a patterned mold by pouring PFPE-DMA containing1-hydroxycyclohexyl phenyl ketone over the patterned area of the master.A poly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the master. The morphology of the mold canthen be confirmed using Atomic Force Microscopy.

Example 5.8. Fabrication of a Perfluoropolyether-Dimethacrylate(PFPE-DMA) Mold from a Template Generated from Carbon Nanotubes

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersing or growing carbonnanotubes on a silicon oxide wafer. This master can be used to templatea patterned mold by pouring PFPE-DMA containing 1-hydroxycyclohexylphenyl ketone over the patterned area of the master. Apoly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the master. The morphology of the mold canthen be confirmed using Atomic Force Microscopy.

Example 6 Method of Making Monodisperse Nanostructures Having aPlurality of Shapes and Sizes

In some embodiments, the presently disclosed subject matter describes anovel “top down” soft lithographic technique; non-wetting imprintlithography (NoWIL) which allows completely isolated nanostructures tobe generated by taking advantage of the inherent low surface energy andswelling resistance of cured PFPE-based materials.

The presently described subject matter provides a novel “top down” softlithographic technique; non-wetting imprint lithography (NoWIL) whichallows completely isolated nanostructures to be generated by takingadvantage of the inherent low surface energy and swelling resistance ofcured PFPE-based materials. Without being bound to any one particulartheory, a key aspect of NoWIL is that both the elastomeric mold and thesurface underneath the drop of monomer or resin are non-wetting to thisdroplet. If the droplet wets this surface, a thin scum layer willinevitably be present even if high pressures are exerted upon the mold.When both the elastomeric mold and the surface are non-wetting (i.e. aPFPE mold and fluorinated surface) the liquid is confined only to thefeatures of the mold and the scum layer is eliminated as a seal formsbetween the elastomeric mold and the surface under a slight pressure.Thus, the presently disclosed subject matter provides for the first timea simple, general, soft lithographic method to produce nanoparticles ofnearly any material, size, and shape that are limited only by theoriginal master used to generate the mold.

Using NoWIL, nanoparticles composed of 3 different polymers weregenerated from a variety of engineered silicon masters. Representativepatterns include, but are not limited to, 3-μm arrows (see FIG. 11 ),conical shapes that are 500 nm at the base and converge to <50 nm at thetip (see FIG. 12 ), and 200-nm trapezoidal structures (see FIG. 13 ).Definitive proof that all particles were indeed “scum-free” wasdemonstrated by the ability to mechanically harvest these particles bysimply pushing a doctor's blade across the surface. See FIGS. 20 and 22.

Polyethylene glycol (PEG) is a material of interest for drug deliveryapplications because it is readily available, non-toxic, andbiocompatible. The use of PEG nanoparticles generated by inversemicroemulsions to be used as gene delivery vectors has previously beenreported. K. McAllister et al., Journal of the American Chemical Society124, 15198-15207 (Dec. 25, 2002). In the presently disclosed subjectmatter, NoWIL was performed using a commercially availablePEG-diacrylate and blending it with 1 wt % of a photoinitiator,1-hydroxycyclohexyl phenyl ketone. PFPE molds were generated from avariety of patterned silicon substrates using a dimethacrylatefunctionalized PFPE oligomer (PFPE DMA) as described previously. See J.P. Rolland, E. C. Hagberg, G. M. Denison, K. R. Carter, J. M. DeSimone,Angewandte Chemie-International Edition 43, 5796-5799 (2004). Flat,uniform, non-wetting surfaces were generated by using a silicon wafertreated with a fluoroalkyl trichlorosilane or by drawing a doctor'sblade across a small drop of PFPE-DMA on a glass substrate andphotocuring. A small drop of PEG diacrylate was then placed on thenon-wetting surface and the patterned PFPE mold placed on top of it. Thesubstrate was then placed in a molding apparatus and a small pressurewas applied to push out the excess PEG-diacrylate. The entire apparatuswas then subjected to UV light (λ=365 nm) for ten minutes while under anitrogen purge. Particles were observed after separation of the PFPEmold and flat, non-wetting substrate using optical microscopy, scanningelectron microscopy (SEM), and atomic force microscopy (AFM).

Poly(lactic acid) (PLA) and derivatives thereof, such aspoly(lactide-co-glycolide) (PLGA), have had a considerable impact on thedrug delivery and medical device communities because it isbiodegradable. See K. E. Uhrich, S. M. Cannizzaro, R. S. Langer, K. M.Shakesheff, Chemical Reviews 99, 3181-3198 (November, 1999); A. C.Albertsson, I. K. Varma, Biomacromolecules 4, 1466-1486(November-December, 2003). As with PEG-based systems, progress has beenmade toward the fabrication of PLGA particles through various dispersiontechniques that result in size distributions and are strictly limited tospherical shapes. See C. Cui, S. P. Schwendeman, Langmuir 34, 8426(2001).

The presently disclosed subject matter demonstrates the use of NoWIL togenerate discrete PLA particles with total control over shape and sizedistribution. For example, in one embodiment, one gram of(3S)-cis-3,6-dimethyl-1,4-dioxane-2,5-dione was heated above its meltingtemperature to 110° C. and ˜20 μL of stannous octoate catalyst/initiatorwas added to the liquid monomer. A drop of the PLA monomer solution wasthen placed into a preheated molding apparatus which contained anon-wetting flat substrate and mold. A small pressure was applied aspreviously described to push out excess PLA monomer. The apparatus wasallowed to heat at 110° C. for 15 h until the polymerization wascomplete. The PFPE-DMA mold and the flat, non-wetting substrate werethen separated to reveal the PLA particles.

To further demonstrate the versatility of NoWIL, particles composed of aconducting polymer polypyrrole (PPy) were generated. PPy particles havebeen formed using dispersion methods, see M. R. Simmons, P. A. Chaloner,S. P. Armes, Langmuir 11, 4222 (1995), as well as “lost-wax” techniques,see P. Jiang, J. F. Bertone, V. L. Colvin, Science 291, 453 (2001).

The presently disclosed subject matter demonstrate for the first time,complete control over shape and size distribution of PPy particles.Pyrrole is known to polymerize instantaneously when in contact withoxidants such as perchloric acid. Dravid et al. has shown that thispolymerization can be retarded by the addition of tetrahydrofuran (THF)to the pyrrole. See M. Su, M. Aslam, L. Fu, N. Q. Wu, V. P. Dravid,Applied Physics Letters 84, 4200-4202 (May 24, 2004).

The presently disclosed subject matter takes advantage of this propertyin the formation of PPy particles by NoWIL. For example, 50 μL of a 1:1v/v solution of THF:pyrrole was added to 50 μL of 70% perchloric acid. Adrop of this clear, brown solution (prior to complete polymerization)into the molding apparatus and applied pressure to remove excesssolution. The apparatus was then placed into the vacuum oven overnightto remove the THF and water. PPy particles were fabricated with goodfidelity using the same masters as previously described.

Importantly, the materials properties and polymerization mechanisms ofPLA, PEG, and PPy are completely different. For example, while PLA is ahigh-modulus, semicrystalline polymer formed using a metal-catalyzedring opening polymerization at high temperature, PEG is a malleable,waxy solid that is photocured free radically, and PPy is a conductingpolymer polymerized using harsh oxidants. The fact that NoWIL can beused to fabricate particles from these diverse classes of polymericmaterials that require very different reaction conditions underscoresits generality and importance.

In addition to its ability to precisely control the size and shape ofparticles, NoWIL offers tremendous opportunities for the facileencapsulation of agents into nanoparticles. As described in Example3-14, NoWIL can be used to encapsulate a 24-mer DNA strand fluorescentlytagged with CY-3 inside the previously described 200 nm trapezoidal PEGparticles. This was accomplished by simply adding the DNA to themonomer/water solution and molding them as described. We were able toconfirm the encapsulation by observing the particles using confocalfluorescence microscopy (see FIG. 28 ). The presently described approachoffers a distinct advantage over other encapsulation methods in that nosurfactants, condensation agents, and the like are required.Furthermore, the fabrication of monodisperse, 200 nm particlescontaining DNA represents a breakthrough step towards artificialviruses. Accordingly, a host of biologically important agents, such asgene fragments, pharmaceuticals, oligonucleotides, and viruses, can beencapsulated by this method.

The method also is amenable to non-biologically oriented agents, such asmetal nanoparticles, crystals, or catalysts. Further, the simplicity ofthis system allows for straightforward adjustment of particleproperties, such as crosslink density, charge, and composition by theaddition of other comonomers, and combinatorial generation of particleformulations that can be tailored for specific applications.

Accordingly, NoWIL is a highly versatile method for the production ofisolated, discrete nanostructures of nearly any size and shape. Theshapes presented herein were engineered non-arbitrary shapes. NoWIL caneasily be used to mold and replicate non-engineered shapes found innature, such as viruses, crystals, proteins, and the like. Furthermore,the technique can generate particles from a wide variety of organic andinorganic materials containing nearly any cargo. The method issimplistically elegant in that it does not involve complex surfactantsor reaction conditions to generate nanoparticles. Finally, the processcan be amplified to an industrial scale by using existing softlithography roller technology, see Y. N. Xia, D. Qin, G. M. Whitesides,Advanced Materials 8, 1015-1017 (December, 1996), or silk screenprinting methods.

Example 7 Synthesis of Functional Perfluoropolyethers Example 7.1.Synthesis of Krytox® (DuPont, Wilmington, Del., United States ofAmerica) Diol to be Used as a Functional PFPE

Example 7.2. Synthesis of Krytox® (DuPont, Wilmington, Del., UnitedStates of America) Diol to be Used as a Functional PFPE

Example 7.3. Synthesis of Krytox® (DuPont, Wilmington, Del., UnitedStates of America) Diol to be Used as a Functional PFPE

Example 7.4. Example of Krytox® (DuPont, Wilmington, Del., United Statesof America) Diol to be Used as a Functional PFPE

Example 7.5. Synthesis of a Multi-Arm PFPE Precursor

wherein, X includes, but is not limited to an isocyanate, an acidchloride, an epoxy, and a halogen; R includes, but is not limited to anacrylate, a methacrylate, a styrene, an epoxy, and an amine; and thecircle represents any multifunctional molecule, such a cyclic compound.PFPE can be any perfluoropolyether material as described herein,including, but not limited to a perfluoropolyether material comprising abackbone structure as follows:

Example 7.6. Synthesis of a Hyperbranched PFPE Precursor

wherein, PFPE can be any perfluoropolyether material as describedherein, including, but not limited to a perfluoropolyether materialcomprising a backbone structure as follows:

It will be understood that various details of the presently disclosedsubject matter can be changed without departing from the scope of thepresently disclosed subject matter. Furthermore, the foregoingdescription is for the purpose of illustration only, and not for thepurpose of limitation.

What is claimed is:
 1. A pharmaceutical composition comprising, aplurality of uniform drug delivery particles prepared by a processcomprising: introducing the pharmaceutical composition into cavities ofa mold, and removing the pharmaceutical composition from the cavities ofthe mold, wherein the pharmaceutical composition retains a physicalshape substantially similar to the cavities, wherein the pharmaceuticalcomposition is present throughout each particle, and wherein theparticles of the plurality of delivery particles have parallel lateraledges and parallel top and bottom edges in cross-section and are lessthan about 100 micrometers in a broadest dimension.
 2. Thepharmaceutical composition of claim 1, wherein the broadest dimension isless than about 10 micrometers.
 3. The pharmaceutical composition ofclaim 1, wherein the broadest dimension is between about 1 micrometerand about 10 micrometers.
 4. The pharmaceutical composition of claim 1,wherein the broadest dimension is between about 1 micrometer and about 3micrometers.
 5. The pharmaceutical composition of claim 1, wherein eachdelivery particle of the plurality is free-standing.
 6. Thepharmaceutical composition of claim 1, wherein the pharmaceuticalcomposition is configured for local delivery.
 7. The pharmaceuticalcomposition of claim 1, wherein the mold comprises a polymer material.8. The pharmaceutical composition of claim 1, wherein the mold comprisesa perfluoropolyether material, an acrylate material, or a siliconematerial.
 9. The pharmaceutical composition of claim 1, wherein theintroducing the pharmaceutical composition into the cavities of the moldcomprises heat or capillary force.
 10. The pharmaceutical composition ofclaim 9, wherein the process further comprises hardening thepharmaceutical composition in the mold cavities prior to the removingthe pharmaceutical composition from the mold cavities.
 11. A pluralityof uniform micro sized drug delivery particles having parallel lateraledges and parallel top and bottom edges in cross-section and apharmaceutical or therapeutic composition present throughout eachparticle, wherein the drug delivery particles are prepared by a processcomprising: introducing the pharmaceutical or therapeutic compositioninto cavities of a mold, and removing the pharmaceutical or therapeuticcomposition from the cavities of the mold, wherein the compositionretains a physical shape substantially similar to the cavity, whereinthe particles have a broadest dimension less than about 100 micrometers.12. The drug delivery particles of claim 11, wherein the particles havea broadest dimension less than about 10 micrometers.
 13. The drugdelivery particles of claim 11, wherein the particles have a broadestdimension between about 1 micrometer and about 10 micrometers.
 14. Thedrug delivery particles of claim 11, wherein the drug delivery particlesare configured for local delivery.
 15. The drug delivery particles ofclaim 11, wherein the mold comprises a polymer material.
 16. The drugdelivery particles of claim 11, wherein the mold comprises aperfluoropolyether material, an acrylate material, or a siliconematerial.
 17. The drug delivery particles of claim 11, wherein theintroducing the pharmaceutical or therapeutic composition into thecavities of the mold comprises heat or capillary force.
 18. The drugdelivery particles of claim 11, wherein the process further compriseshardening the pharmaceutical or therapeutic composition in the moldcavities prior to the removing the pharmaceutical or therapeuticcomposition from the mold cavities.
 19. A method of treating a subjectwith uniform micro sized drug delivery particles having parallel lateraledges and parallel top and bottom edges in cross-section and apharmaceutical or therapeutic composition present throughout eachparticle, said method comprising, administering the uniform drugdelivery particles to a patient in need thereof, wherein the drugdelivery particles are prepared by a process comprising: introducing thepharmaceutical or therapeutic composition into cavities of a mold, andremoving the pharmaceutical or therapeutic composition from the cavitiesof the mold, wherein the composition retains a physical shapesubstantially similar to the cavity, wherein the particles have abroadest dimension less than about 100 micrometers.